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Open AccessReview

Application of Chitosan in Bone and Dental Engineering

INSERM (French National Institute of Health and Medical Research), UMR 1260, Regenerative Nanomedicine (RNM), FMTS, 11 Rue Humann, 67000 Strasbourg, France
Université de Strasbourg, Faculté de Chirurgie Dentaire de Strasbourg, 8 Rue Sainte-Elisabeth, 67000 Strasbourg, France
Hôpitaux Universitaires de Strasbourg, Pôle de Médecine et de Chirurgie Bucco-Dentaires, 67000 Strasbourg, France
Authors to whom correspondence should be addressed.
Academic Editor: Jyh-Ping Chen
Molecules 2019, 24(16), 3009;
Received: 14 July 2019 / Revised: 8 August 2019 / Accepted: 19 August 2019 / Published: 19 August 2019
(This article belongs to the Special Issue Chitosan-Based Nanomaterials for Biomedical Applications)


Chitosan is a deacetylated polysaccharide from chitin, the natural biopolymer primarily found in shells of marine crustaceans and fungi cell walls. Upon deacetylation, the protonation of free amino groups of the d-glucosamine residues of chitosan turns it into a polycation, which can easily interact with DNA, proteins, lipids, or negatively charged synthetic polymers. This positive-charged characteristic of chitosan not only increases its solubility, biodegradability, and biocompatibility, but also directly contributes to the muco-adhesion, hemostasis, and antimicrobial properties of chitosan. Combined with its low-cost and economic nature, chitosan has been extensively studied and widely used in biopharmaceutical and biomedical applications for several decades. In this review, we summarize the current chitosan-based applications for bone and dental engineering. Combining chitosan-based scaffolds with other nature or synthetic polymers and biomaterials induces their mechanical properties and bioactivities, as well as promoting osteogenesis. Incorporating the bioactive molecules into these biocomposite scaffolds accelerates new bone regeneration and enhances neovascularization in vivo.
Keywords: chitosan; bone engineering; regeneration; scaffold; periodontitis; dental pulp chitosan; bone engineering; regeneration; scaffold; periodontitis; dental pulp

1. Introduction

The first bioerodable artificial polymer-cell scaffold was implanted into animals 30 years ago [1]. Since then, tissue engineering has become an interdisciplinary field that applies the principles of engineering and life sciences toward the development of biological substitutes that restore, maintain, or improve tissue structure and function [2]. Several biomaterials have been used for the fabrication of the scaffolds, including natural materials derived from animals or plants (collagen, starch, gelatin, alginate, cellulose, fibrin, hyaluronan, and chitosan) and synthetic materials, such as bioactive ceramics and a wide range of synthetic polymers. However, the excellent bio-based 3-dimentional (3D) polymer scaffolds should not only be non-toxic, biocompatible, and biodegradable, but also be competent in promoting cell adhesion and retaining the metabolic functions of attached cells [3], as the 3D polymer scaffolds used in tissue engineering should mimic and provide an actual in vivo surrounding microenvironment for the incorporation of cells or growth factors to regenerate damaged tissues or organs [4]. The immunomodulatory biological effects of chitosan-based scaffold have also been described. In this regard, chitosan becomes one of the most commonly studied polymers in the scientific research, not only for biopharmaceutical and biomedical applications, but also for food science and technology [5].

2. Results

2.1. Chitosan

Chitosan (CS) is a linear, semi-crystalline polysaccharide composed of β-(1→4)-2-acetamido-2-deoxy-b-d-glucan (n-acetyl d-glucosamine) and β-(1→4)-2-amino-2-deoxyb-d-glucan (d-glucosamine) units [6,7]. Its molecular weight ranges from 10 to over 1000 kDa. Chitosan is not extensively present in the environment, however, it can be easily produced via the alkaline n-deacetylation process of the natural biopolymer commonly found in the shells of marine crustaceans and in fungi cells walls—the chitin [8,9] (Figure 1). The deacetylation degree (DD) of chitosan, which gives an indication of the number of amino groups along the chains, is calculated as the ratio of d-glucosamine to the sum of d-glucosamine and n-acetyl d-glucosamine [7,10,11].
Chitin is a white, hard, inelastic, nitrogeneous polysaccharide. It is hydrophobic and is not soluble in water and most organic solvents, except for hexafluroisopropanol, hexafluroacetone, and chloroalcohols [12]. This poor solubility of chitin is an extreme limit for its practical applications. However, the free amino groups of the d-glucosamine residues of chitosan, which could be protonated, provide a better solubility for chitosan by forming a non-Newtonian, shear-thinning fluid in most diluted acidic solutions at a pH below 6.5 (pKa value ~6.3) [7,13] (Figure 1). With protonated amino groups, chitosan becomes a polycation and could subsequently form ionic complexes with a wide variety of natural or synthetic anionic species, for example, DNA, proteins, lipids, or negatively charged synthetic polymers such as poly(acrylic acid) [7,14,15].
Chitosan can be biodegraded into non-toxic residues by lysozyme or chitinase, which hydrolyses glucosamine-glucosamine, glucosamine-n-acetyl-glucosamine, and n-acetyl-glucosamine-n-acetyl-glucosamine linkages [16,17,18]. The rate and extent of chitosan’s biodegradability in living organisms are highly related to the molecular mass of the polymer and its deacetylation degree (DD) [19,20]. The chitosan DD can also influence its biocompatibility. A higher DD increases the number of positive charges which increases the interaction between chitosan and cells, leading to an improved biocompatibility [21]. In addition, chitosan is a low-cost and economic natural biopolymer [22]. The price of chitin (3.6–6.0 US$/kg)/chitosan (30–500 US$/kg) are hundreds or thousands of times higher than the price of shell wastes (0.05–0.15 US$/kg) [23], while the production costs were around 1.70 US$/kg for chitin and 3.50 US$/kg for chitosan 40 years ago [24].

2.2. Medical and Pharmaceutical Properties of Chitosan

As a natural multifunctional polysaccharide, chitosan has been widely studied for biomedical, surgical, and tissue engineering and pharmaceutical application, thanks to its biocompatibility, biodegradability, and muco-adhesiveness. Chitosan is reported to be increasingly used in the United States as an over-the-counter cholesterol-lowering agent. Positively-charged deacetylated chitosan could bind negatively charged molecules, such as fatty acids, lipids, and bile acids, in the intestinal tract, excreting these molecules from the body [25]. Chitosan is thus considered as a promising candidate for obesity and hypercholesterolemia treatment [26]. Chitosan could be also used for wastewater treatment and beverage clarification because of its good chelating or binding capacity of protonable amino groups for various species, such as metal ions [27,28]. However, the most important medical and pharmaceutical applications of chitosan are drug delivery, wound dressings, and biocomposite scaffolds for tissue engineering.

2.2.1. Drug Delivery

The positively charged protonable amino group of the d-glucosamine residues of chitosan can interact with the negatively charged sialic acid residues of the glycoprotein which composes the mucus. Thus, the muco-adhesion is directly related to the DD of chitosan, with higher chitosan DD resulting in increased positive charges which enhance its muco-adhesive properties [29]. It has also been reported that chitosan can interact with the negative part of the cell membrane, enhancing the penetration of an active agent through the epithelium layer that contains tight junctions [30,31]. Consequently, owing to the muco-adhesion and enhanced penetration properties, chitosan is a suitable excipient to prepare oral, nasal, ocular, vaginal, and subcutaneous delivery forms and be used as a vaccine adjuvant or co-adjuvant to enhance the bioavailability and immunogenicity of antigens [32,33,34].

2.2.2. Wound Dressings

Wound healing is a particular biological phenomenon, which progresses through a series of inter-reliant and corresponding stages to regenerate the integrity of damaged tissue and replacement of lost tissue [35]. The hemostatic and antimicrobial properties of chitosan enable its application in wound dressings [35,36,37].
Again, as a natural positive-charged polysaccharide, protonable amino groups on the chitosan backbone electrostatically interact with the various negatively charged proteins and glycolipids on the surface of red blood cells (RBC). This interaction increases blood viscosity, activates platelet adhesion and aggregation, and enhances the transportation of platelets to the vascular wall for physiological hemostasis. Blood clots are formed by intensive aggregation of RBC around the wound site to quickly stop bleeding [38,39,40,41]. Thus, the number of positive charged amino groups on chitosan directly plays an important role in its hemostatic property.
The antimicrobial activity of chitosan has been demonstrated against different microorganisms both in vivo and in vitro, such as bacteria (either Gram-positive or Gram-negative), yeast, fungi, and algae, which makes chitosan a good candidate as an antimicrobial agent in solution, film, and composite [35,42,43,44,45,46]. This antimicrobial property of chitosan has been related to the presence of its cationic nature. However, this antimicrobial activity could also depend on other intrinsic factors (for example, the type of chitosan or the degree of chitosan polymerization and deacetylation) and extrinsic factors (such as the live host, the microorganisms, and the environmental conditions). Until now, the antimicrobial activity of chitosan has not yet been fully understood.

2.3. Chitosan-Based Scaffold Preparation

The most common method to generate chitosan scaffolds is by freezing and lyophilizing chitosan solution. The spaces occupied by ice crystals formed in frozen chitosan solution are emptied during the sublimation, leading to the formation of pores. However, a precise control of temperature is required to form good pore structures [47,48]. Another method to form porous scaffolds is called salt leaching [49,50]. Salt crystals such as NaCl are used as porogens and put into a mold, and chitosan is then poured over the salt, penetrating into all the small spaces left between the salt crystals. The mold is then heated to melt the chitosan powder in an oven for a sufficient time. The chilled chitosan/NaCl mixture is then separated from the mold and the salt is washed away by water or alcohol, generating open pores in the chitosan scaffolds. In addition, fibrous chitosan scaffolds are formed by electrospinning. An applied electric field causes the elongation of the chitosan drop and leads to the formation of long fibers ranging from the submicron level to several nanometers in diameter [51,52].
Besides the dried chitosan-based scaffold preparation discussed above, chitosan-based scaffolds could also be prepared as hydrogels [53]. A hydrogel is a network of the same or different types of cross-linked polymer chains with good water absorption capacity. Chemically cross-linked hydrogels are formed by covalent linking of chitosan macromers, in which the bond formation is irreversible. Covalent cross-linking can also be formed between polymers and a cross-linker [54], or via photopolymerization [55]. Another way to prepare chitosan-based hydrogel is by physical cross-linking through ionic interactions. Polyelectrolyte complex networks are formed via ionic interactions between positively charged chitosan and anions or other negatively charged polymers. Although chemically cross-linked chitosan-based hydrogels show better stability and resistance to environmental variables, physically cross-linked hydrogels are more biocompatible because of the lack of chemical cross-linkers.

2.4. Chitosan-Based Scaffolds for Bone Regeneration

Bone is a hard and highly functionalized connective tissue constituting the skeletal framework of the human body. It supports fleshy structures, protects vital organs, and is involved in various physiological functions such as the maintenance of phosphocalcic homeostasis [56,57]. After an injury, if the impairment is mild and the defect size is small, bone tissue has certain self-healing potential through osteogenic differentiation of bone marrow mesenchymal stem cells, bone neoformation, and neo-angiogenesis at the lesion site. For large-size bone defects, bone grafts are usually needed [58,59], however, these clinical procedures have many disadvantages [60].
In the context of bone engineering, the physiological inertness and low toxic effects of chitosan-based scaffolds have been demonstrated by numerous in vitro and in vivo studies (Table 1). In addition, no allergic and inflammatory reactions upon chitosan-based material implantation, injection, or topical application in the human body have been proven [61,62,63]; however, the use of chitosan as scaffold is limited by having reduced bioactivities and mechanical properties. This disadvantage has been overcome by mixing the chitosan scaffolds with other synthetic or natural polymers [poly(vinyl alcohol), poly-ɛ-caprolactone, alginate, collagen, silk fibroin, etc.], biomaterials (hydroxyapatite, β-tricalcium phosphate, SiO2, etc.), or bioactive pharmacological molecules (bone morphogenetic protein 2 (BMP-2), vascular endothelial growth factor (VEGF), bisphosphonate, etc.).
Indeed, blending chitosan with most of the aforementioned polymer and/or biomaterials efficiently reinforces its mechanical properties and improves its bioactivities, such as increased protein absorption and increased biomineralization (Table 1). However, many of the published biocomposite scaffolds could not decrease the biodegradation rate of chitosan, which affects their long-term in vivo persistence (Table 1). Water retention ability is another parameter important for tissue engineering scaffolds, as the increased water retention ability of scaffold implanted in vivo could lead to the loosening and dislocation of the implant. As summarized in Table 1, most of the biocomposite scaffolds reported controls and decreased their water retention ability.
Most importantly, both in vitro and in vivo experiments demonstrated that all these chitosan-based biocomposite scaffolds are not toxic, and have very good properties of biocompatibility, osteoconductivity, and osteogenesis, promoting good cell proliferation and cell adhesion, causing an increase in new bone regeneration. Regarding more specific clinical applications of chitosan in alveolar bone and jawbone regeneration, several preclinical reports showed promising results, even for accelerating dental implant osseointegration and reconstructing critical size defects [64,65,66,67].
Over the last decade, the functionality of these chitosan-based biocomposite scaffolds have been enhanced by incorporating bioactive molecules into drug delivery systems. Therefore, bioactive molecules could be locally delivered with an adequate dose for a desired period, avoiding the drug release to non-target sites. Bone morphogenetic proteins (BMPs) are a group of growth factors originally discovered by their ability to induce bone and cartilage formation [68]. Transforming growth factor beta 1 (TGF-β1) is a polypeptide member of the transforming growth factor beta superfamily of cytokines, controlling cell growth, cell proliferation, and cell differentiation in many cell types. Studies have shown that chitosan-based biocomposite associated with either BMP2, BMP-7 or TGF-β1 efficiently induced osteogenesis and promoted a large amount new bone formation compared with control or sham groups (see Table 1). Vascular endothelial growth factor (VEGF) is a signal protein stimulating the formation of blood vessels. Incorporating VEGF into chitosan-based biocomposite scaffolds induces angiogenesis and enhances neovascularization in bone healing (see Table 1).

2.5. Chitosan-Based Application for Dental Engineering: The Case of Periodontal Regeneration

Periodontitis is a chronic inflammatory disease induced by bacterial infection, affecting tooth-supporting tissues and leading to significant destruction of the periodontium (i.e., gingiva, alveolar bone, periodontal ligament, cement). Such a disease is common in the global population (>50% in the U.S. population) and is the main cause of tooth loss [69]. It is characterized by the development of the periodontal pocket and, in severe cases, of infrabony defects. Treatment of such lesions represent a challenge for the clinician [70,71]. Indeed, periodontal treatment aims to reduce the inflammation and to control the infection by chemical (antibiotics, antiseptics) and mechanical treatments (scaling and root planing). However, such a treatment strategy is associated mainly to wound repair, characterized by a long junctional epithelium. In most severe cases (probing pocket depth (PPD) > 5 mm), non-surgical treatment alone could be insufficient in emphasizing the need of adjuvant therapy [72,73]. Therefore, the use of local delivery of active drugs or compounds has been suggested to target inflammation and infection, and in promoting tissue regeneration [74,75,76,77]. During the last decade, a specific interest was made towards the chitosan-based delivery system [78]. Several chitosan-based devices have been designed and evaluated in specific contexts, including in micro/nanoparticles, fibers, membrane, and gels [79,80,81]. Chitosan gels exhibit interesting physical properties that could be modulated by the concentration of chitosan. Indeed, it was shown that such chitosan-based gels (1–4%) have an interesting viscosity to be injected within periodontal pockets. Most importantly, they can be used as a reliable vehicle to release active drugs at the disease site. While a sustained release could be considered ideal, it is important to note that the kinetic of release is a parameter that is also influenced by the percentage of chitosan [82]. Drugs, such as statins [83], doxycycline [84], or other antibiotics/antiseptics such as tetracyclines, have been incorporated in such devices [85,86,87].
Interestingly, adjuvant use of such drug-loaded scaffolds improved periodontal healing, emphasizing the interest in their use [78]. For instance, Chang et al. evaluated in vivo, in an experimental periodontitis murine model, the use of a chitosan-based hydrogel loaded with naringin, a natural compound with anti-inflammatory properties. They demonstrated the interesting properties of chitosan hydrogel, as its injection at the lesion site was associated with a burst release of the active compound during the acute phase of the inflammation, inducing an anti-inflammatory effect within periodontal tissues [88]. Such results might be explained by the controlled local delivery at the lesion site and the 3D characteristics of the chitosan scaffold.
Interestingly, it was also demonstrated that chitosan can potentialize the antibacterial effect of chlorhexidine [89], showing the intrinsic property of chitosan.

2.6. Chitosan-Based Scaffold in Dental-Pulp Regeneration

The endodontic space located in the heart of the tooth contains pulp, which is crucial for its vitality. This soft connective tissue contains collagen fibers, dental stem cells, vascularization, and innervation surrounded by dentin. This pulp-dentin complex is very reactive and, after significant aggression, the painful inflammatory reaction of the pulp–dentin complex is often hard to dissipate and leads to pulp necrosis. The conventional endodontic treatment is drastic, as all tissues are removed whatever their regenerative potentials. Recently, regenerative strategies targeting regeneration of pulp connective-tissue, dentin, radicular edification, vascularization, and innervation were developed [143]. Scaffolds represent the key element of the endodontic treatment. They are essential for delivering active molecules and for carrying competent cells within the endodontic compartment. They must have an adequate viscosity, allowing their injection in the whole root canal system, and porosity, which is crucial for cell colonization.
Recently, chitosan-based scaffolds have been developed for this purpose [144]. They are favorable, not only for pulp regeneration, but also for dentin formation because of their capacities to induce mineralization. Indeed, chitosan scaffolds containing β-tricalcium phosphate promoted a high expression of mineralization markers, such as osteopontin and alkaline phosphatase, and dentin formation by human periodontal ligament cells (HPLCs) [145]. In addition, several recent reports demonstrated that chitosan-based scaffolds promoted the proliferation, migration, and odontoblastic differentiation of dental pulp stem cells and mesenchymal stem cells both in vitro and in vivo [146,147,148,149] (Table 2). The use of chitosan as a vector for antimicrobials has also been tested with interesting outcomes [150].

3. Conclusions

As a positive-charged, low-cost natural polymer with good biodegradability and biocompatibility, as well as having non-toxic, muco-adhesive, hemostatic, and antimicrobial properties, chitosan is a good candidate for biomedical and biopharmaceutical research. Consequently, chitosan-based scaffolds have been widely studied and applied in tissue engineering for the last two decades. Indeed, blending chitosan with other natural or synthetic polymers and/or biomaterials could efficiently control the porosity and water retention of these biocomposite scaffolds, reduce their biodegradation rate, enhance their bioactivity and biocompatibility, and increase their mechanical properties. Importantly, as demonstrated in both in vitro and in vivo studies, most of these biocomposite scaffolds have no cytotoxicity and promote the attachment and proliferation of cells for tissue repair. In addition, the combination of bioactive molecules with these chitosan-based biocomposite scaffolds could not only promote the proliferation and differentiation of stem cells and accelerate the tissue regeneration, but also induce angiogenesis and vascularization in different animal models, which could consequently be used in human clinical trials.

Author Contributions

A.A., F.B., F.C., F.F., O.H., N.B.-J., and G.H. conceived and designed the review; A.A., N.Z., E.H., B.H., F.B., D.O., F.C., F.F., O.H., N.B.-J., and G.H. studied the literature; A.A., N.Z., F.B., D.O., F.C., F.F., O.H., N.B.-J., and G.H. wrote the manuscript.


This research was supported by the Agence Nationale de la Recherche (ANR), Institut National de la Santé et de la Recherche Médicale (INSERM), and Faculté de Chirurgie-Dentaire (Université de Strasbourg).

Conflicts of Interest

The authors declare no conflict of interest.


  1. Vacanti, J.P.; Morse, M.A.; Saltzman, W.M.; Domb, A.J.; Perez-Atayde, A.; Langer, R. Selective cell transplantation using bioabsorbable artificial polymers as matrices. J. Pediatr. Surg. 1988, 23, 3–9. [Google Scholar] [CrossRef]
  2. Langer, R.; Vacanti, J.P. Tissue engineering. Science 1993, 260, 920–926. [Google Scholar] [CrossRef] [PubMed]
  3. Chan, B.P.; Leong, K.W. Scaffolding in tissue engineering: General approaches and tissue-specific considerations. Eur. Spine. J. 2008, 17, 467–479. [Google Scholar] [CrossRef] [PubMed]
  4. Loh, Q.L.; Choong, C. Three-Dimensional Scaffolds for Tissue Engineering Applications: Role of Porosity and Pore Size. Tissue Eng. Part B Rev. 2013, 19, 485–502. [Google Scholar] [CrossRef] [PubMed]
  5. Bellich, B.; D’Agostino, I.; Semeraro, S.; Gamini, A.; Cesàro, A. “The Good, the Bad and the Ugly” of Chitosans. Mar. Drugs 2016, 14, 99. [Google Scholar] [CrossRef] [PubMed]
  6. Deepthi, S.; Venkatesan, J.; Kim, S.-K.; Bumgardner, J.D.; Jayakumar, R. An overview of chitin or chitosan/nano ceramic composite scaffolds for bone tissue engineering. Int. J. Biol. Macromol. 2016, 93, 1338–1353. [Google Scholar] [CrossRef] [PubMed]
  7. Croisier, F.; Jérôme, C. Chitosan-based biomaterials for tissue engineering. Eur. Polym. J. 2013, 49, 780–792. [Google Scholar] [CrossRef]
  8. Aranaz, I.; Harris, R.; Heras, A. Chitosan Amphiphilic Derivatives. Chem. Appl. Curr. Org. Chem. 2010, 308–330. [Google Scholar] [CrossRef]
  9. Dash, M.; Chiellini, F.; Ottenbrite, R.M.; Chiellini, E. Chitosan—A versatile semi-synthetic polymer in biomedical applications. Prog. Polym. Sci. 2011, 36, 981–1014. [Google Scholar] [CrossRef]
  10. Madihally, S.V.; Matthew, H.W. Porous chitosan scaffolds for tissue engineering. Biomaterials 1999, 20, 1133–1142. [Google Scholar] [CrossRef]
  11. Acosta, N.; Jiménez, C.; Borau, V.; Heras, A. Extraction and characterization of chitin from crustaceans. Biomass Bioenergy 1993, 5, 145–153. [Google Scholar] [CrossRef]
  12. Synowiecki, J.; Al-Khateeb, N.A. Production, properties, and some new applications of chitin and its derivatives. Crit. Rev. Food Sci. Nutr. 2003, 43, 145–171. [Google Scholar] [CrossRef] [PubMed]
  13. Yi, H.; Wu, L.-Q.; Bentley, W.E.; Ghodssi, R.; Rubloff, G.W.; Culver, J.N.; Payne, G.F. Biofabrication with chitosan. Biomacromolecules 2005, 6, 2881–2894. [Google Scholar] [CrossRef] [PubMed]
  14. Pavinatto, F.J.; Caseli, L.; Oliveira, O.N. Chitosan in nanostructured thin films. Biomacromolecules 2010, 11, 1897–1908. [Google Scholar] [CrossRef] [PubMed]
  15. Takahashi, T.; Takayama, K.; Machida, Y.; Nagai, T. Characteristics of polyion complexes of chitosan with sodium alginate and sodium polyacrylate. Int. J. Pharm. 1990, 61, 35–41. [Google Scholar] [CrossRef]
  16. Kean, T.; Thanou, M. Biodegradation, biodistribution and toxicity of chitosan. Adv. Drug Deliv. Rev. 2010, 62, 3–11. [Google Scholar] [CrossRef] [PubMed]
  17. Baldrick, P. The safety of chitosan as a pharmaceutical excipient. Regul. Toxicol. Pharm. 2010, 56, 290–299. [Google Scholar] [CrossRef] [PubMed]
  18. Huet, J.; Rucktooa, P.; Clantin, B.; Azarkan, M.; Looze, Y.; Villeret, V.; Wintjens, R. X-ray structure of papaya chitinase reveals the substrate binding mode of glycosyl hydrolase family 19 chitinases. Biochemistry 2008, 47, 8283–8291. [Google Scholar] [CrossRef] [PubMed]
  19. Yang, Y.M.; Hu, W.; Wang, X.D.; Gu, X.S. The controlling biodegradation of chitosan fibers by N-acetylation in vitro and in vivo. J. Mater. Sci Mater. Med. 2007, 18, 2117–2121. [Google Scholar] [CrossRef]
  20. Xu, J.; McCarthy, S.P.; Gross, R.A.; Kaplan, D.L. Chitosan Film Acylation and Effects on Biodegradability. Macromolecules 1996, 29, 3436–3440. [Google Scholar] [CrossRef]
  21. Chatelet, C.; Damour, O.; Domard, A. Influence of the degree of acetylation on some biological properties of chitosan films. Biomaterials 2001, 22, 261–268. [Google Scholar] [CrossRef]
  22. Kadouche, S.; Farhat, M.; Lounici, H.; Fiallo, M.; Sharrock, P.; Mecherri, M.; Hadioui, M. Low Cost Chitosan Biopolymer for Environmental Use Made from Abundant Shrimp Wastes. Waste Biomass Valor. 2017, 8, 401–406. [Google Scholar] [CrossRef]
  23. Pichyangkura, R. Application of chitin-chitosan from marine by-products in Thailand. In The FFTC-KU Joint Seminar on “Improved Utilization of Fishery By-product as Potential Nutraceuticals and Functional Foods”; Kasetsart University: Bangkok, Thailand, 2014; pp. 190–194. [Google Scholar]
  24. Roberts, G.A.F. Thirty years of progress in chitin and chitosan. Prog. Chem. Appl. Chitin. Its Deriv. 2018, 13, 7–15. [Google Scholar]
  25. Tapola, N.S.; Lyyra, M.L.; Kolehmainen, R.M.; Sarkkinen, E.S.; Schauss, A.G. Safety aspects and cholesterol-lowering efficacy of chitosan tablets. J. Am. Coll. Nutr. 2008, 27, 22–30. [Google Scholar] [CrossRef] [PubMed]
  26. Baker, W.L.; Tercius, A.; Anglade, M.; White, C.M.; Coleman, C.I. A meta-analysis evaluating the impact of chitosan on serum lipids in hypercholesterolemic patients. Ann. Nutr. Metab. 2009, 55, 368–374. [Google Scholar] [CrossRef] [PubMed]
  27. Rytwo, G.; Lavi, R.; Rytwo, Y.; Monchase, H.; Dultz, S.; König, T.N. Clarification of olive mill and winery wastewater by means of clay-polymer nanocomposites. Sci. Total Env. 2013, 442, 134–142. [Google Scholar] [CrossRef] [PubMed]
  28. Rytwo, G. The use of clay-polymer nanocomposites in wastewater pretreatment. Scientific World J. 2012, 2012, 498503. [Google Scholar] [CrossRef]
  29. He, P.; Davis, S.S.; Illum, L. In vitro evaluation of the mucoadhesive properties of chitosan microspheres. Int. J. Pharm. 1998, 166, 75–88. [Google Scholar] [CrossRef]
  30. Yeh, T.-H.; Hsu, L.-W.; Tseng, M.T.; Lee, P.-L.; Sonjae, K.; Ho, Y.-C.; Sung, H.-W. Mechanism and consequence of chitosan-mediated reversible epithelial tight junction opening. Biomaterials 2011, 32, 6164–6173. [Google Scholar] [CrossRef]
  31. Smith, J.; Wood, E.; Dornish, M. Effect of chitosan on epithelial cell tight junctions. Pharm. Res. 2004, 21, 43–49. [Google Scholar] [CrossRef]
  32. Van der Lubben, I.M.; Kersten, G.; Fretz, M.M.; Beuvery, C.; Coos Verhoef, J.; Junginger, H.E. Chitosan microparticles for mucosal vaccination against diphtheria: Oral and nasal efficacy studies in mice. Vaccine 2003, 21, 1400–1408. [Google Scholar] [CrossRef]
  33. Slütter, B.; Bal, S.M.; Que, I.; Kaijzel, E.; Löwik, C.; Bouwstra, J.; Jiskoot, W. Antigen-adjuvant nanoconjugates for nasal vaccination: An improvement over the use of nanoparticles? Mol. Pharm. 2010, 7, 2207–2215. [Google Scholar] [CrossRef]
  34. Bal, S.M.; Slütter, B.; Verheul, R.; Bouwstra, J.A.; Jiskoot, W. Adjuvanted, antigen loaded N-trimethyl chitosan nanoparticles for nasal and intradermal vaccination: Adjuvant-and site-dependent immunogenicity in mice. Eur. J. Pharm. Sci. 2012, 45, 475–481. [Google Scholar] [CrossRef]
  35. Boateng, J.S.; Matthews, K.H.; Stevens, H.N.E.; Eccleston, G.M. Wound healing dressings and drug delivery systems: A review. J. Pharm. Sci. 2008, 97, 2892–2923. [Google Scholar] [CrossRef]
  36. Hurler, J.; Skalko-Basnet, N. Potentials of chitosan-based delivery systems in wound therapy: Bioadhesion study. J. Funct. Biomater. 2012, 3, 37–48. [Google Scholar] [CrossRef]
  37. Okamoto, Y.; Yano, R.; Miyatake, K.; Tomohiro, I.; Shigemasa, Y.; Minami, S. Effects of chitin and chitosan on blood coagulation. Carbohydr. Polym. 2003, 53, 337–342. [Google Scholar] [CrossRef]
  38. Hu, Z.; Zhang, D.-Y.; Lu, S.-T.; Li, P.-W.; Li, S.-D. Chitosan-Based Composite Materials for Prospective Hemostatic Applications. Mar. Drugs 2018, 16, 273. [Google Scholar] [CrossRef]
  39. Huang, Y.; Feng, L.; Zhang, Y.; He, L.; Wang, C.; Xu, J.; Wu, J.; Kirk, T.B.; Guo, R.; Xue, W. Hemostasis mechanism and applications of N-alkylated chitosan sponge. Polym. Adv. Technol. 2017, 28, 1107–1114. [Google Scholar] [CrossRef]
  40. Chou, T.-C.; Fu, E.; Wu, C.-J.; Yeh, J.-H. Chitosan enhances platelet adhesion and aggregation. Biochem. Biophys. Res. Commun. 2003, 302, 480–483. [Google Scholar] [CrossRef]
  41. Sagnella, S.; Mai-Ngam, K. Chitosan based surfactant polymers designed to improve blood compatibility on biomaterials. Colloids Surf. B Biointerfaces 2005, 42, 147–155. [Google Scholar] [CrossRef]
  42. Ahmed, S.; Ikram, S. Chitosan Based Scaffolds and Their Applications in Wound Healing. Achiev. Life Sci. 2016, 10, 27–37. [Google Scholar] [CrossRef]
  43. Choi, B.K.; Kim, K.Y.; Yoo, Y.J.; Oh, S.J.; Choi, J.H.; Kim, C.Y. In vitro antimicrobial activity of a chitooligosaccharide mixture against Actinobacillus actinomycetemcomitans and Streptococcus mutans. Int. J. Antimicrob. Agents 2001, 18, 553–557. [Google Scholar] [CrossRef]
  44. Ikinci, G.; Senel, S.; Akincibay, H.; Kaş, S.; Erciş, S.; Wilson, C.G.; Hincal, A.A. Effect of chitosan on a periodontal pathogen Porphyromonas gingivalis. Int. J. Pharm. 2002, 235, 121–127. [Google Scholar] [CrossRef]
  45. Savard, T.; Beaulieu, C.; Boucher, I.; Champagne, C.P. Antimicrobial action of hydrolyzed chitosan against spoilage yeasts and lactic acid bacteria of fermented vegetables. J. Food Prot. 2002, 65, 828–833. [Google Scholar] [CrossRef]
  46. Martínez-Camacho, A.P.; Cortez-Rocha, M.O.; Ezquerra-Brauer, J.M.; Graciano-Verdugo, A.Z.; Rodriguez-Félix, F.; Castillo-Ortega, M.M.; Yépiz-Gómez, M.S.; Plascencia-Jatomea, M. Chitosan composite films: Thermal, structural, mechanical and antifungal properties. Carbohydr. Polym. 2010, 82, 305–315. [Google Scholar] [CrossRef]
  47. Ruiz-Caro, R.; Veiga-Ochoa, M.D. Characterization and dissolution study of chitosan freeze-dried systems for drug controlled release. Molecules 2009, 14, 4370–4386. [Google Scholar] [CrossRef]
  48. Keller, L.; Regiel-Futyra, A.; Gimeno, M.; Eap, S.; Mendoza, G.; Andreu, V.; Wagner, Q.; Kyzioł, A.; Sebastian, V.; Stochel, G.; et al. Chitosan-based nanocomposites for the repair of bone defects. Nanomedicine 2017, 13, 2231–2240. [Google Scholar] [CrossRef]
  49. Lim, J.I.; Lee, Y.-K.; Shin, J.-S.; Lim, K.-J. Preparation of interconnected porous chitosan scaffolds by sodium acetate particulate leaching. J. Biomater. Sci. Polym. Ed. 2011, 22, 1319–1329. [Google Scholar] [CrossRef]
  50. Pezeshki-Modaress, M.; Rajabi-Zeleti, S.; Zandi, M.; Mirzadeh, H.; Sodeifi, N.; Nekookar, A.; Aghdami, N. Cell-loaded gelatin/chitosan scaffolds fabricated by salt-leaching/lyophilization for skin tissue engineering: In vitro and in vivo study. J. Biomed. Mater. Res. A 2014, 102, 3908–3917. [Google Scholar] [CrossRef]
  51. Geng, X.; Kwon, O.-H.; Jang, J. Electrospinning of chitosan dissolved in concentrated acetic acid solution. Biomaterials 2005, 26, 5427–5432. [Google Scholar] [CrossRef]
  52. Dahlin, R.L.; Kasper, F.K.; Mikos, A.G. Polymeric nanofibers in tissue engineering. Tissue Eng. Part. B Rev. 2011, 17, 349–364. [Google Scholar] [CrossRef]
  53. Ahmadi, F.; Oveisi, Z.; Samani, S.M.; Amoozgar, Z. Chitosan based hydrogels: Characteristics and pharmaceutical applications. Res. Pharm. Sci. 2015, 10, 1–16. [Google Scholar]
  54. Bhattarai, N.; Gunn, J.; Zhang, M. Chitosan-based hydrogels for controlled, localized drug delivery. Adv. Drug Deliv. Rev. 2010, 62, 83–99. [Google Scholar] [CrossRef]
  55. Chen, R.; Chen, Q.; Huo, D.; Ding, Y.; Hu, Y.; Jiang, X. In situ formation of chitosan-gold hybrid hydrogel and its application for drug delivery. Colloids Surf B Biointerfaces 2012, 97, 132–137. [Google Scholar] [CrossRef]
  56. Walsh, J.S. Normal bone physiology, remodelling and its hormonal regulation. Surg. (Oxf) 2015, 33, 1–6. [Google Scholar] [CrossRef]
  57. Arvidson, K.; Abdallah, B.M.; Applegate, L.A.; Baldini, N.; Cenni, E.; Gomez-Barrena, E.; Granchi, D.; Kassem, M.; Konttinen, Y.T.; Mustafa, K.; et al. Bone regeneration and stem cells. J. Cell Mol. Med. 2011, 15, 718–746. [Google Scholar] [CrossRef]
  58. Loi, F.; Córdova, L.A.; Pajarinen, J.; Lin, T.; Yao, Z.; Goodman, S.B. Inflammation, fracture and bone repair. Bone 2016, 86, 119–130. [Google Scholar] [CrossRef]
  59. Gómez-Barrena, E.; Rosset, P.; Lozano, D.; Stanovici, J.; Ermthaller, C.; Gerbhard, F. Bone fracture healing: Cell therapy in delayed unions and nonunions. Bone 2015, 70, 93–101. [Google Scholar] [CrossRef]
  60. Rogers, G.F.; Greene, A.K. Autogenous bone graft: basic science and clinical implications. J. Craniofac. Surg. 2012, 23, 323–327. [Google Scholar] [CrossRef]
  61. Waibel, K.H.; Haney, B.; Moore, M.; Whisman, B.; Gomez, R. Safety of chitosan bandages in shellfish allergic patients. Mil. Med. 2011, 176, 1153–1156. [Google Scholar] [CrossRef]
  62. Jiang, T.; Ji, H.; Zhang, L.; Wang, Y.; Zhou, H. Chitosan Oligosaccharide Exerts Anti-Allergic Effect against Shrimp Tropomyosin-Induced Food Allergy by Affecting Th1 and Th2 Cytokines. Int. Arch. Allergy Immunol. 2019, 1–7. [Google Scholar] [CrossRef]
  63. Azuma, K.; Osaki, T.; Minami, S.; Okamoto, Y. Anticancer and anti-inflammatory properties of chitin and chitosan oligosaccharides. J. Funct. Biomater. 2015, 6, 33–49. [Google Scholar] [CrossRef]
  64. Bhattarai, G.; Lee, Y.-H.; Lee, M.-H.; Park, I.-S.; Yi, H.-K. Insulin-like growth factor binding protein-3 affects osteogenic efficacy on dental implants in rat mandible. Mater. Sci. Eng. C Mater. Biol. Appl. 2015, 55, 490–496. [Google Scholar] [CrossRef]
  65. Bhattarai, G.; Lee, Y.H.; Lee, M.H.; Yi, H.K. Gene delivery of c-myb increases bone formation surrounding oral implants. J. Dent. Res. 2013, 92, 840–845. [Google Scholar] [CrossRef]
  66. Lee, Y.-H.; Kim, J.-S.; Kim, J.-E.; Lee, M.-H.; Jeon, J.-G.; Park, I.-S.; Yi, H.-K. Nanoparticle mediated PPARγ gene delivery on dental implants improves osseointegration via mitochondrial biogenesis in diabetes mellitus rat model. Nanomedicine 2017, 13, 1821–1832. [Google Scholar] [CrossRef]
  67. Li, H.; Ji, Q.; Chen, X.; Sun, Y.; Xu, Q.; Deng, P.; Hu, F.; Yang, J. Accelerated bony defect healing based on chitosan thermosensitive hydrogel scaffolds embedded with chitosan nanoparticles for the delivery of BMP2 plasmid DNA. J. Biomed. Mater. Res. A 2017, 105, 265–273. [Google Scholar] [CrossRef]
  68. Bleuming, S.A.; He, X.C.; Kodach, L.L.; Hardwick, J.C.; Koopman, F.A.; ten Kate, F.J.; van Deventer, S.J.H.; Hommes, D.W.; Peppelenbosch, M.P.; Offerhaus, G.J.; et al. Bone Morphogenetic Protein Signaling Suppresses Tumorigenesis at Gastric Epithelial Transition Zones in Mice. Cancer Res. 2007, 67, 8149–8155. [Google Scholar] [CrossRef]
  69. Kinane, D.F.; Stathopoulou, P.G.; Papapanou, P.N. Periodontal diseases. Nat. Rev. Dis. Primers 2017, 3, 17038. [Google Scholar] [CrossRef]
  70. Cortellini, P.; Tonetti, M.S. Clinical concepts for regenerative therapy in intrabony defects. Periodontol. 2000 2015, 68, 282–307. [Google Scholar] [CrossRef]
  71. Cortellini, P.; Tonetti, M.S. Focus on intrabony defects: Guided tissue regeneration. Periodontol. 2000 2000, 22, 104–132. [Google Scholar] [CrossRef]
  72. Harmouche, L.; Courval, A.; Mathieu, A.; Petit, C.; Huck, O.; Severac, F.; Davideau, J.-L. Impact of tooth-related factors on photodynamic therapy effectiveness during active periodontal therapy: A 6-months split-mouth randomized clinical trial. Photodiagnosis Photodyn. 2019, 190–194. [Google Scholar] [CrossRef]
  73. Abdul Rahman, N.; Nickles, K.; Gallenbach, K.; Dannewitz, B.; Ramich, T.; Scharf, S.; Röllke, L.; Schacher, B.; Eickholz, P. Five-year stability of clinical attachment after regenerative treatment of infrabony defects compared to controls. J. Clin. Periodontol. 2019, 46, 650–658. [Google Scholar] [CrossRef]
  74. Da Rocha, H.A.J.; Silva, C.F.; Santiago, F.L.; Martins, L.G.; Dias, P.C.; De Magalhães, D. Local Drug Delivery Systems in the Treatment of Periodontitis: A Literature Review. J. Int. Acad. Periodontol. 2015, 17, 82–90. [Google Scholar]
  75. Morand, D.N.; Davideau, J.-L.; Clauss, F.; Jessel, N.; Tenenbaum, H.; Huck, O. Cytokines during periodontal wound healing: Potential application for new therapeutic approach. Oral Dis. 2017, 23, 300–311. [Google Scholar] [CrossRef]
  76. Batool, F.; Strub, M.; Petit, C.; Bugueno, I.M.; Bornert, F.; Clauss, F.; Huck, O.; Kuchler-Bopp, S.; Benkirane-Jessel, N. Periodontal Tissues, Maxillary Jaw Bone, and Tooth Regeneration Approaches: From Animal Models Analyses to Clinical Applications. Nanomaterials 2018, 8, 337. [Google Scholar] [CrossRef]
  77. Petit, C.; Batool, F.; Bugueno, I.M.; Schwinté, P.; Benkirane-Jessel, N.; Huck, O. Contribution of Statins towards Periodontal Treatment: A Review. Mediat. Inflamm. 2019, 2019, 6367402. [Google Scholar] [CrossRef]
  78. Sah, A.K.; Dewangan, M.; Suresh, P.K. Potential of chitosan-based carrier for periodontal drug delivery. Colloids Surf. B: Biointerfaces 2019, 178, 185–198. [Google Scholar] [CrossRef]
  79. Yar, M.; Farooq, A.; Shahzadi, L.; Khan, A.S.; Mahmood, N.; Rauf, A.; Chaudhry, A.A.; Rehman, I.U. Novel meloxicam releasing electrospun polymer/ceramic reinforced biodegradable membranes for periodontal regeneration applications. Mater. Sci. Eng. C Mater. Biol. Appl. 2016, 64, 148–156. [Google Scholar] [CrossRef]
  80. Kamal, T.; Khan, S.B.; Haider, S.; Alghamdi, Y.G.; Asiri, A.M. Thin layer chitosan-coated cellulose filter paper as substrate for immobilization of catalytic cobalt nanoparticles. Int. J. Biol. Macromol. 2017, 104, 56–62. [Google Scholar] [CrossRef]
  81. Işılay Özdoğan, A.; Akca, G.; Şenel, S. Development and in vitro evaluation of chitosan based system for local delivery of atorvastatin for treatment of periodontitis. Eur. J. Pharm. Sci. 2018, 124, 208–216. [Google Scholar] [CrossRef]
  82. Ganguly, A.; Ian, C.K.; Sheshala, R.; Sahu, P.S.; Al-Waeli, H.; Meka, V.S. Application of diverse natural polymers in the design of oral gels for the treatment of periodontal diseases. J. Mater. Sci. Mater. Med. 2017, 28, 39. [Google Scholar] [CrossRef]
  83. Özdoğan, A.I.; İlarslan, Y.D.; Kösemehmetoğlu, K.; Akca, G.; Kutlu, H.B.; Comerdov, E.; Iskit, A.B.; Şenel, S. In vivo evaluation of chitosan based local delivery systems for atorvastatin in treatment of periodontitis. Int. J. Pharm. 2018, 550, 470–476. [Google Scholar] [CrossRef]
  84. Madi, M.; Pavlic, V.; Samy, W.; Alagl, A. The anti-inflammatory effect of locally delivered nano-doxycycline gel in therapy of chronic periodontitis. Acta Odontol. Scand. 2018, 76, 71–76. [Google Scholar] [CrossRef]
  85. Khan, G.; Yadav, S.K.; Patel, R.R.; Nath, G.; Bansal, M.; Mishra, B. Development and Evaluation of Biodegradable Chitosan Films of Metronidazole and Levofloxacin for the Management of Periodontitis. Aaps Pharmscitech 2016, 17, 1312–1325. [Google Scholar] [CrossRef]
  86. Lin, C.-Y.; Tseng, K.-S.; Liu, J.-M.; Chuang, H.-C.; Lien, C.-H.; Chen, Y.-C.; Lai, C.-Y.; Yu, C.-P.; Hsu, R.-J. Increased Risk of Ulcerative Colitis in Patients with Periodontal Disease: A Nationwide Population-Based Cohort Study. Int. J. Env. Res. Public Health 2018, 15, 2602. [Google Scholar] [CrossRef]
  87. Kilicarslan, M.; Gumustas, M.; Yildiz, S.; Baykara, T. Preparation and characterization of chitosan-based spray-dried microparticles for the delivery of clindamycin phosphate to periodontal pockets. Curr. Drug Deliv. 2014, 11, 98–111. [Google Scholar] [CrossRef]
  88. Chang, P.-C.; Chao, Y.-C.; Hsiao, M.-H.; Chou, H.-S.; Jheng, Y.-H.; Yu, X.-H.; Lee, N.; Yang, C.; Liu, D.-M. Inhibition of Periodontitis Induction Using a Stimuli-Responsive Hydrogel Carrying Naringin. J. Periodontol. 2017, 88, 190–196. [Google Scholar] [CrossRef]
  89. Barreras, U.S.; Méndez, F.T.; Martínez, R.E.M.; Valencia, C.S.; Rodríguez, P.R.M.; Rodríguez, J.P.L. Chitosan nanoparticles enhance the antibacterial activity of chlorhexidine in collagen membranes used for periapical guided tissue regeneration. Mater. Sci. Eng. C Mater. Biol. Appl. 2016, 58, 1182–1187. [Google Scholar] [CrossRef]
  90. Ferrand, A.; Eap, S.; Richert, L.; Lemoine, S.; Kalaskar, D.; Demoustier-Champagne, S.; Atmani, H.; Mély, Y.; Fioretti, F.; Schlatter, G.; et al. Osteogenetic properties of electrospun nanofibrous PCL scaffolds equipped with chitosan-based nanoreservoirs of growth factors. Macromol. Biosci. 2014, 14, 45–55. [Google Scholar] [CrossRef]
  91. Issa, J.P.M.; do Nascimento, C.; Bentley, M.V.L.B.; Del Bel, E.A.; Iyomasa, M.M.; Sebald, W.; de Albuquerque, R.F. Bone repair in rat mandible by rhBMP-2 associated with two carriers. Micron 2008, 39, 373–379. [Google Scholar] [CrossRef]
  92. Seol, Y.-J.; Lee, J.-Y.; Park, Y.-J.; Lee, Y.-M.; Young-Ku, N.; Rhyu, I.-C.; Lee, S.-J.; Han, S.-B.; Chung, C.-P. Chitosan sponges as tissue engineering scaffolds for bone formation. Biotechnol. Lett. 2004, 26, 1037–1041. [Google Scholar] [CrossRef]
  93. Mohammadi, R.; Amini, K. Guided bone regeneration of mandibles using chitosan scaffold seeded with characterized uncultured omental adipose-derived stromal vascular fraction: An animal study. Int. J. Oral Maxillofac Implant. 2015, 30, 216–222. [Google Scholar] [CrossRef]
  94. Xu, F.; Wu, Y.; Zhang, Y.; Yin, P.; Fang, C.; Wang, J. Influence of in vitro differentiation status on the in vivo bone regeneration of cell/chitosan microspheres using a rat cranial defect model. J. Biomater. Sci. Polym. Ed. 2019, 30, 1008–1025. [Google Scholar] [CrossRef]
  95. Danilchenko, S.N.; Kalinkevich, O.V.; Pogorelov, M.V.; Kalinkevich, A.N.; Sklyar, A.M.; Kalinichenko, T.G.; Ilyashenko, V.Y.; Starikov, V.V.; Bumeyster, V.I.; Sikora, V.Z.; et al. Characterization and in vivo evaluation of chitosan-hydroxyapatite bone scaffolds made by one step coprecipitation method. J. Biomed. Mater. Res. Part. A 2011, 96A, 639–647. [Google Scholar] [CrossRef]
  96. Zhang, X.; Zhu, L.; Lv, H.; Cao, Y.; Liu, Y.; Xu, Y.; Ye, W.; Wang, J. Repair of rabbit femoral condyle bone defects with injectable nanohydroxyapatite/chitosan composites. J. Mater. Sci. Mater. Med. 2012, 23, 1941–1949. [Google Scholar] [CrossRef]
  97. Gaihre, B.; Jayasuriya, A.C. Comparative investigation of porous nano-hydroxyapaptite/chitosan, nano-zirconia/chitosan and novel nano-calcium zirconate/chitosan composite scaffolds for their potential applications in bone regeneration. Mater. Sci. Eng. C Mater. Biol. Appl. 2018, 91, 330–339. [Google Scholar] [CrossRef]
  98. Cho, B.C.; Chung, H.Y.; Lee, D.G.; Yang, J.D.; Park, J.W.; Roh, K.H.; Kim, G.U.; Lee, D.S.; Kwon, I.C.; Bae, E.H.; et al. The effect of chitosan bead encapsulating calcium sulfate as an injectable bone substitute on consolidation in the mandibular distraction osteogenesis of a dog model. J. Oral Maxillofac. Surg. 2005, 63, 1753–1764. [Google Scholar] [CrossRef]
  99. Pattnaik, S.; Nethala, S.; Tripathi, A.; Saravanan, S.; Moorthi, A.; Selvamurugan, N. Chitosan scaffolds containing silicon dioxide and zirconia nano particles for bone tissue engineering. Int. J. Biol. Macromol. 2011, 49, 1167–1172. [Google Scholar] [CrossRef]
  100. Shokri, S.; Movahedi, B.; Rafieinia, M.; Salehi, H. A new approach to fabrication of Cs/BG/CNT nanocomposite scaffold towards bone tissue engineering and evaluation of its properties. Appl. Surf. Sci. 2015, 357, 1758–1764. [Google Scholar] [CrossRef]
  101. Kucharska, M.; Butruk, B.; Walenko, K.; Brynk, T.; Ciach, T. Fabrication of in-situ foamed chitosan/β-TCP scaffolds for bone tissue engineering application. Mater. Lett. 2012, 85, 124–127. [Google Scholar] [CrossRef]
  102. Cheng, G.; Li, Z.; Wan, Q.; Lv, K.; Li, D.; Xing, X.; Li, Z. A novel animal model treated with tooth extraction to repair the full-thickness defects in the mandible of rabbits. J. Surg. Res. 2015, 194, 706–716. [Google Scholar] [CrossRef]
  103. Chang, H.-H.; Wang, Y.-L.; Chiang, Y.-C.; Chen, Y.-L.; Chuang, Y.-H.; Tsai, S.-J.; Heish, K.-H.; Lin, F.-H.; Lin, C.-P. A Novel Chitosan-γPGA Polyelectrolyte Complex Hydrogel Promotes Early New Bone Formation in the Alveolar Socket Following Tooth Extraction. PloS ONE 2014, 9, e92362. [Google Scholar] [CrossRef]
  104. Fan, J.; Park, H.; Lee, M.K.; Bezouglaia, O.; Fartash, A.; Kim, J.; Aghaloo, T.; Lee, M. Adipose-derived stem cells and BMP-2 delivery in chitosan-based 3D constructs to enhance bone regeneration in a rat mandibular defect model. Tissue Eng. Part. A 2014, 20, 2169–2179. [Google Scholar] [CrossRef]
  105. Pourhaghgouy, M.; Zamanian, A.; Shahrezaee, M.; Masouleh, M.P. Physicochemical properties and bioactivity of freeze-cast chitosan nanocomposite scaffolds reinforced with bioactive glass. Mater. Sci. Eng. C Mater. Biol. Appl. 2016, 58, 180–186. [Google Scholar] [CrossRef]
  106. Nazemi, K.; Azadpour, P.; Moztarzadeh, F.; Urbanska, A.M.; Mozafari, M. Tissue-engineered chitosan/bioactive glass bone scaffolds integrated with PLGA nanoparticles: A therapeutic design for on-demand drug delivery. Mater. Lett. 2015, 138, 16–20. [Google Scholar] [CrossRef]
  107. Venkatesan, J.; Ryu, B.; Sudha, P.N.; Kim, S.-K. Preparation and characterization of chitosan–carbon nanotube scaffolds for bone tissue engineering. Int. J. Biol. Macromol. 2012, 50, 393–402. [Google Scholar] [CrossRef]
  108. Saravanan, S.; Sameera, D.K.; Moorthi, A.; Selvamurugan, N. Chitosan scaffolds containing chicken feather keratin nanoparticles for bone tissue engineering. Int. J. Biol. Macromol. 2013, 62, 481–486. [Google Scholar] [CrossRef]
  109. Maglione, M.; Spano, S.; Ruaro, M.E.; Salvador, E.; Zanconati, F.; Tromba, G.; Turco, G. In vivo evaluation of chitosan-glycerol gel scaffolds seeded with stem cells for full-thickness mandibular bone regeneration. J. Oral Sci. 2017, 59, 225–232. [Google Scholar] [CrossRef]
  110. Saravanan, S.; Vimalraj, S.; Anuradha, D. Chitosan based thermoresponsive hydrogel containing graphene oxide for bone tissue repair. Biomed. Pharm. 2018, 107, 908–917. [Google Scholar] [CrossRef]
  111. Keller, L.; Pijnenburg, L.; Idoux-Gillet, Y.; Bornert, F.; Benameur, L.; Tabrizian, M.; Auvray, P.; Rosset, P.; Gonzalo-Daganzo, R.M.; Barrena, E.G.; et al. Preclinical safety study of a combined therapeutic bone wound dressing for osteoarticular regeneration. Nat. Commun. 2019, 10, 2156. [Google Scholar] [CrossRef]
  112. Jayakumar, R.; Ramachandran, R.; Sudheesh Kumar, P.T.; Divyarani, V.V.; Srinivasan, S.; Chennazhi, K.P.; Tamura, H.; Nair, S.V. Fabrication of chitin–chitosan/nano ZrO2 composite scaffolds for tissue engineering applications. Int. J. Biol. Macromol. 2011, 49, 274–280. [Google Scholar] [CrossRef]
  113. Park, J.-S.; Choi, S.-H.; Moon, I.-S.; Cho, K.-S.; Chai, J.-K.; Kim, C.-K. Eight-week histological analysis on the effect of chitosan on surgically created one-wall intrabony defects in beagle dogs. J. Clin. Periodontol. 2003, 30, 443–453. [Google Scholar] [CrossRef]
  114. Eap, S.; Ferrand, A.; Schiavi, J.; Keller, L.; Kokten, T.; Fioretti, F.; Mainard, D.; Ladam, G.; Benkirane-Jessel, N. Collagen implants equipped with ’fish scale’-like nanoreservoirs of growth factors for bone regeneration. Nanomed. (Lond) 2014, 9, 1253–1261. [Google Scholar] [CrossRef]
  115. Zhang, Y.; Shi, B.; Li, C.; Wang, Y.; Chen, Y.; Zhang, W.; Luo, T.; Cheng, X. The synergetic bone-forming effects of combinations of growth factors expressed by adenovirus vectors on chitosan/collagen scaffolds. J. Control. Release 2009, 136, 172–178. [Google Scholar] [CrossRef]
  116. Zhang, Q.; Hubenak, J.; Iyyanki, T.; Alred, E.; Turza, K.C.; Davis, G.; Chang, E.I.; Branch-Brooks, C.D.; Beahm, E.K.; Butler, C.E. Engineering vascularized soft tissue flaps in an animal model using human adipose-derived stem cells and VEGF+PLGA/PEG microspheres on a collagen-chitosan scaffold with a flow-through vascular pedicle. Biomaterials 2015, 73, 198–213. [Google Scholar] [CrossRef]
  117. Shi, S.; Cheng, X.; Wang, J.; Zhang, W.; Peng, L.; Zhang, Y. RhBMP-2 microspheres-loaded chitosan/collagen scaffold enhanced osseointegration: An experiment in dog. J. Biomater. Appl. 2009, 23, 331–346. [Google Scholar]
  118. Elango, J.; Saravanakumar, K.; Rahman, S.U.; Henrotin, Y.; Regenstein, J.M.; Wu, W.; Bao, B. Chitosan-Collagen 3D Matrix Mimics Trabecular Bone and Regulates RANKL-Mediated Paracrine Cues of Differentiated Osteoblast and Mesenchymal Stem Cells for Bone Marrow Macrophage-Derived Osteoclastogenesis. Biomolecules 2019, 9, 173. [Google Scholar] [CrossRef]
  119. Ansarizadeh, M.; Mashayekhan, S.; Saadatmand, M. Fabrication, modeling and optimization of lyophilized advanced platelet rich fibrin in combination with collagen-chitosan as a guided bone regeneration membrane. Int. J. Biol. Macromol. 2019, 125, 383–391. [Google Scholar] [CrossRef]
  120. Sowjanya, J.A.; Singh, J.; Mohita, T.; Sarvanan, S.; Moorthi, A.; Srinivasan, N.; Selvamurugan, N. Biocomposite scaffolds containing chitosan/alginate/nano-silica for bone tissue engineering. Colloids Surf. B Biointerfaces 2013, 109, 294–300. [Google Scholar] [CrossRef]
  121. Kim, H.-L.; Jung, G.-Y.; Yoon, J.-H.; Han, J.-S.; Park, Y.-J.; Kim, D.-G.; Zhang, M.; Kim, D.-J. Preparation and characterization of nano-sized hydroxyapatite/alginate/chitosan composite scaffolds for bone tissue engineering. Mater. Sci. Eng. C Mater. Biol. Appl. 2015, 54, 20–25. [Google Scholar] [CrossRef]
  122. Jin, H.-H.; Kim, D.-H.; Kim, T.-W.; Shin, K.-K.; Jung, J.S.; Park, H.-C.; Yoon, S.-Y. In vivo evaluation of porous hydroxyapatite/chitosan–alginate composite scaffolds for bone tissue engineering. Int. J. Biol. Macromol. 2012, 51, 1079–1085. [Google Scholar] [CrossRef]
  123. Florczyk, S.J.; Leung, M.; Li, Z.; Huang, J.I.; Hopper, R.A.; Zhang, M. Evaluation of three-dimensional porous chitosan-alginate scaffolds in rat calvarial defects for bone regeneration applications. J. Biomed. Mater. Res. A 2013, 101, 2974–2983. [Google Scholar] [CrossRef]
  124. Teimouri, A.; Ebrahimi, R.; Emadi, R.; Beni, B.H.; Chermahini, A.N. Nano-composite of silk fibroin-chitosan/Nano ZrO2 for tissue engineering applications: Fabrication and morphology. Int. J. Biol. Macromol. 2015, 76, 292–302. [Google Scholar] [CrossRef]
  125. Lima, P.A.L.; Resende, C.X.; de Almeida Soares, G.D.; Anselme, K.; Almeida, L.E. Preparation, characterization and biological test of 3D-scaffolds based on chitosan, fibroin and hydroxyapatite for bone tissue engineering. Mater. Sci. Eng. C Mater. Biol. Appl. 2013, 33, 3389–3395. [Google Scholar] [CrossRef]
  126. Tong, S.; Xu, D.-P.; Liu, Z.-M.; Du, Y.; Wang, X.-K. Synthesis of and in vitro and in vivo evaluation of a novel TGF-β1-SF-CS three-dimensional scaffold for bone tissue engineering. Int. J. Mol. Med. 2016, 38, 367–380. [Google Scholar] [CrossRef]
  127. Niu, X.; Fan, Y.; Liu, X.; Li, X.; Li, P.; Wang, J.; Sha, Z.; Feng, Q. Repair of bone defect in femoral condyle using microencapsulated chitosan, nanohydroxyapatite/collagen and poly(L-lactide)-based microsphere-scaffold delivery system. Artif. Organs 2011, 35, E119–E128. [Google Scholar] [CrossRef]
  128. Sainitya, R.; Sriram, M.; Kalyanaraman, V.; Dhivya, S.; Saravanan, S.; Vairamani, M.; Sastry, T.P.; Selvamurugan, N. Scaffolds containing chitosan/carboxymethyl cellulose/mesoporous wollastonite for bone tissue engineering. Int. J. Biol. Macromol. 2015, 80, 481–488. [Google Scholar] [CrossRef]
  129. Miranda, S.C.C.C.; Silva, G.A.B.; Mendes, R.M.; Abreu, F.A.M.; Caliari, M.V.; Alves, J.B.; Goes, A.M. Mesenchymal stem cells associated with porous chitosan–gelatin scaffold: A potential strategy for alveolar bone regeneration. J. Biomed. Mater. Res. Part. A 2012, 100A, 2775–2786. [Google Scholar] [CrossRef]
  130. Olad, A.; Farshi Azhar, F. The synergetic effect of bioactive ceramic and nanoclay on the properties of chitosan–gelatin/nanohydroxyapatite–montmorillonite scaffold for bone tissue engineering. Ceram. Int. 2014, 40, 10061–10072. [Google Scholar] [CrossRef]
  131. Kavya, K.C.; Jayakumar, R.; Nair, S.; Chennazhi, K.P. Fabrication and characterization of chitosan/gelatin/nSiO2 composite scaffold for bone tissue engineering. Int. J. Biol. Macromol. 2013, 59, 255–263. [Google Scholar] [CrossRef]
  132. Serra, I.R.; Fradique, R.; Vallejo, M.C.S.; Correia, T.R.; Miguel, S.P.; Correia, I.J. Production and characterization of chitosan/gelatin/β-TCP scaffolds for improved bone tissue regeneration. Mater. Sci. Eng. C Mater. Biol. Appl. 2015, 55, 592–604. [Google Scholar] [CrossRef]
  133. Acevedo, C.A.; Olguín, Y.; Briceño, M.; Forero, J.C.; Osses, N.; Díaz-Calderón, P.; Jaques, A.; Ortiz, R. Design of a biodegradable UV-irradiated gelatin-chitosan/nanocomposed membrane with osteogenic ability for application in bone regeneration. Mater. Sci. Eng. C Mater. Biol. Appl. 2019, 99, 875–886. [Google Scholar] [CrossRef]
  134. Maji, K.; Dasgupta, S.; Kundu, B.; Bissoyi, A. Development of gelatin-chitosan-hydroxyapatite based bioactive bone scaffold with controlled pore size and mechanical strength. J. Biomater. Sci. Polym. Ed. 2015, 26, 1190–1209. [Google Scholar] [CrossRef]
  135. Puvaneswary, S.; Talebian, S.; Raghavendran, H.B.; Murali, M.R.; Mehrali, M.; Afifi, A.M.; Kasim, N.H.B.A.; Kamarul, T. Fabrication and in vitro biological activity of βTCP-Chitosan-Fucoidan composite for bone tissue engineering. Carbohydr. Polym. 2015, 134, 799–807. [Google Scholar] [CrossRef]
  136. Jing, X.; Mi, H.-Y.; Peng, J.; Peng, X.-F.; Turng, L.-S. Electrospun aligned poly(propylene carbonate) microfibers with chitosan nanofibers as tissue engineering scaffolds. Carbohydr. Polym. 2015, 117, 941–949. [Google Scholar] [CrossRef]
  137. Zhang, S.; Prabhakaran, M.P.; Qin, X.; Ramakrishna, S. Biocomposite scaffolds for bone regeneration: Role of chitosan and hydroxyapatite within poly-3-hydroxybutyrate-co-3-hydroxyvalerate on mechanical properties and in vitro evaluation. J. Mech. Behav. Biomed. Mater. 2015, 51, 88–98. [Google Scholar] [CrossRef]
  138. Yao, Q.; Li, W.; Yu, S.; Ma, L.; Jin, D.; Boccaccini, A.R.; Liu, Y. Multifunctional chitosan/polyvinyl pyrrolidone/45S5 Bioglass® scaffolds for MC3T3-E1 cell stimulation and drug release. Mater. Sci. Eng. C Mater. Biol. Appl. 2015, 56, 473–480. [Google Scholar] [CrossRef]
  139. Sajesh, K.M.; Jayakumar, R.; Nair, S.V.; Chennazhi, K.P. Biocompatible conducting chitosan/polypyrrole-alginate composite scaffold for bone tissue engineering. Int. J. Biol. Macromol. 2013, 62, 465–471. [Google Scholar] [CrossRef]
  140. Pon-On, W.; Charoenphandhu, N.; Teerapornpuntakit, J.; Thongbunchoo, J.; Krishnamra, N.; Tang, I.-M. Mechanical properties, biological activity and protein controlled release by poly(vinyl alcohol)-bioglass/chitosan-collagen composite scaffolds: A bone tissue engineering applications. Mater. Sci. Eng. C Mater. Biol. Appl. 2014, 38, 63–72. [Google Scholar] [CrossRef]
  141. Peng, L.; Zhou, Y.; Lu, W.; Zhu, W.; Li, Y.; Chen, K.; Zhang, G.; Xu, J.; Deng, Z.; Wang, D. Characterization of a novel polyvinyl alcohol/chitosan porous hydrogel combined with bone marrow mesenchymal stem cells and its application in articular cartilage repair. BMC Musculoskelet. Disord. 2019, 20, 257. [Google Scholar] [CrossRef]
  142. De la Riva, B.; Nowak, C.; Sánchez, E.; Hernández, A.; Schulz-Siegmund, M.; Pec, M.K.; Delgado, A.; Evora, C. VEGF-controlled release within a bone defect from alginate/chitosan/PLA-H scaffolds. Eur. J. Pharm. Biopharm. 2009, 73, 50–58. [Google Scholar] [CrossRef]
  143. Keller, L.; Offner, D.; Schwinté, P.; Morand, D.; Wagner, Q.; Gros, C.; Bornert, F.; Bahi, S.; Musset, A.-M.; Benkirane-Jessel, N.; et al. Active Nanomaterials to Meet the Challenge of Dental Pulp Regeneration. Mater. (Basel) 2015, 8, 7461–7471. [Google Scholar] [CrossRef]
  144. Zein, N.; Harmouch, E.; Lutz, J.-C.; Fernandez De Grado, G.; Kuchler-Bopp, S.; Clauss, F.; Offner, D.; Hua, G.; Benkirane-Jessel, N.; Fioretti, F. Polymer-Based Instructive Scaffolds for Endodontic Regeneration. Mater. (Basel) 2019, 12, 2347. [Google Scholar] [CrossRef]
  145. Liao, F.; Chen, Y.; Li, Z.; Wang, Y.; Shi, B.; Gong, Z.; Cheng, X. A novel bioactive three-dimensional beta-tricalcium phosphate/chitosan scaffold for periodontal tissue engineering. J. Mater. Sci. Mater. Med. 2010, 21, 489–496. [Google Scholar] [CrossRef]
  146. Albuquerque, M.T.P.; Valera, M.C.; Nakashima, M.; Nör, J.E.; Bottino, M.C. Tissue-engineering-based strategies for regenerative endodontics. J. Dent. Res. 2014, 93, 1222–1231. [Google Scholar] [CrossRef]
  147. Bordini, E.A.F.; Cassiano, F.B.; Silva, I.S.P.; Usberti, F.R.; Anovazzi, G.; Pacheco, L.E.; Pansani, T.N.; Leite, M.L.; Hebling, J.; de Souza Costa, C.A.; et al. Synergistic potential of 1α,25-dihydroxyvitamin D3 and calcium-aluminate-chitosan scaffolds with dental pulp cells. Clin. Oral Investig. 2019, 1–12. [Google Scholar] [CrossRef]
  148. Ducret, M.; Montembault, A.; Josse, J.; Pasdeloup, M.; Celle, A.; Benchrih, R.; Mallein-Gerin, F.; Alliot-Licht, B.; David, L.; Farges, J.-C. Design and characterization of a chitosan-enriched fibrin hydrogel for human dental pulp regeneration. Dent. Mater. 2019, 35, 523–533. [Google Scholar] [CrossRef]
  149. Zhu, N.; Chatzistavrou, X.; Ge, L.; Qin, M.; Papagerakis, P.; Wang, Y. Biological properties of modified bioactive glass on dental pulp cells. J. Dent. 2019, 83, 18–26. [Google Scholar] [CrossRef]
  150. Loyola-Rodríguez, J.P.; Torres-Méndez, F.; Espinosa-Cristobal, L.F.; García-Cortes, J.O.; Loyola-Leyva, A.; González, F.J.; Soto-Barreras, U.; Nieto-Aguilar, R.; Contreras-Palma, G. Antimicrobial activity of endodontic sealers and medications containing chitosan and silver nanoparticles against Enterococcus faecalis. J. Appl. Biomater. Funct. Mater. 2019, 17, 2280800019851771. [Google Scholar] [CrossRef]
Figure 1. Structures for chitin, the noncharged form of chitosan, and protonated positive-charged chitosan.
Figure 1. Structures for chitin, the noncharged form of chitosan, and protonated positive-charged chitosan.
Molecules 24 03009 g001
Table 1. In vitro and in vivo studies of chitosan-based biocomposite scaffolds in bone engineering. VEGF = vascular endothelial growth factor, TGF-β1 = transforming growth factor beta 1.
Table 1. In vitro and in vivo studies of chitosan-based biocomposite scaffolds in bone engineering. VEGF = vascular endothelial growth factor, TGF-β1 = transforming growth factor beta 1.
Chitosan-Based Biocomposite Scaffolds
Polymers and/or BiomaterialsBioactive MoleculeModelsObservations as Compared to the Properties of Chitosan Polymer Scaffolds AloneReference
BMP-2In vitroNo cytotoxicity and increased osteogenesis[90]
In vivoNo cytotoxicity, increased biomineralization, and increased osteogenesis
Recombinant human BMP-2In vivoEnhanced bone regeneration[91]
In vitroIncreased biomineralization and increased osteogenesis[92]
In vivoImproved and earlier bone regeneration[93]
In vitroNo cytotoxicity, increased biomineralization[94]
In vivoGeneration of a substantial amount of bone in rat cranium
Hydroxyapatite In vivoNew bone tissue formation in rat[95]
Nano hydroxyapatite In vivoRegeneration of segmental bone defects with cortical bone in rabbit[96]
Nano hydroxyapatite/Nano ZrO2/Nano CaZrO3 In vitroNo cytotoxicity, decreased water retention and increased mechanical properties[97]
Calcium sulfate In vivoEarly bony consolidation[98]
SiO2 + ZrO2 In vitroNo cytotoxicity at low concentration, decreased water retention, increased protein adsorption, biomineralization, and biodegradation[99]
Bioactive glass + carbon nanotube In vitroNo cytotoxicity, increased water retention, biodegradation, and mechanical properties[100]
β-tricalcium phosphate In vitroNo cytotoxicity at low concentration, decreased biodegradation, and increased mechanical properties[101]
In vivoIncreased new bone formation[102]
γ-polyglutamic acid In vivoIncreased new bone formation[103]
Chondroitine sulfate + apatiteBMP-2In vivoEnhanced bone regeneration[104]
Bioactive glass In vitroDecreased water retention, increased biomineralization, biodegradation, and mechanical properties [105]
Bioactive glass + poly lactic-co-glycolic acid (PLGA) nanoparticles In vitroDecreased water retention, and increased mechanical properties [106]
Carbon nanotube In vitroNo cytotoxicity, increased biomineralization[107]
Keratin nanoparticles In vitroNo cytotoxicity, increased protein adsorption and biodegradation[108]
Glycerophosphate In vivoEnhanced bone regeneration[109]
Glycerophosphate + graphene oxide In vitroNo cytotoxicity, increased water retention, protein adsorption, biomineralization, biodegradation, and osteogenesis[110]
poly-ɛ-caprolactoneBMP-2In vitroNo cytotoxicity[111]
In vivoRegeneration of both the subchondral bone and the cartilage in large animal model
Chitin + Nano ZrO2 In vitroNo cytotoxicity, decreased water retention and biodegradation, increased biomineralization and osteogenesis[112]
Collagen In vivoEnhanced bone regeneration[113]
BMP-2In vitroNo cytotoxicity, increased biomineralization and osteogenesis[114]
In vivoNo cytotoxicity, increased biomineralization and osteogenesis
BMP-7In vivoAccelerated regeneration of alveolar bone tissue[115]
PLGA/Polyethylene glycol (PEG)VEGFIn vitroInduced angiogenesis[116]
In vivoInduced angiogenesis and vascularization in rat
PLGArhBMP-2In vitroControlled growth factor release rate[117]
In vivoEnhanced bone formation and fast bone regeneration in dog
Chondroitine sulfate + hydroxyapatite In vitroSecretion of higher level of receptor activator of nuclear factor kappa-B ligand (RANKL) to mediate osteoclastogenesis[118]
Advanced platelet rich fibrin (A-PRF) In vitroNo cytotoxicity, increased biomineralization and mechanical properties, decreased biodegradation[119]
AlginateNano SiO2 In vitroNo cytotoxicity, decreased water retention and mechanical properties, increased protein adsorption, biomineralization, biodegradation, and osteogenesis[120]
Nano-sized hydroxyapatite In vitroNo cytotoxicity, increased biomineralization, osteogenesis, and mechanical properties[121]
Hydroxyapatite In vitroNo cytotoxicity[122]
In vivoStrong positive effect on bone formation in mice
BMP-2In vitroNo cytotoxicity[123]
In vivoGreat osteogenesis and reconstruction of critical size bone defects
Silk fibroinNano ZrO2 In vitroNo cytotoxicity, increased water retention, biomineralization, biodegradation, and mechanical properties[124]
Hydroxyapatite In vitroIncreased biomineralization and osteogenesis[125]
TGF-β1In vivoBiocompatibility and extensive osteoconductivity and osteogenesis[126]
Collagen +
NanohydroxyapatiteBMP-2In vitroControlled growth factor release rate and more favorable cytocompatibility[127]
In vivoAccelerated regeneration of cancellous bone defect in rabbit
Carboxyme-thylcellulosemesoporous wollastonite In vitroNo cytotoxicity, decreased water retention and biodegradation, increased protein adsorption, biomineralization, and osteogenesis[128]
Gelatin In vivoIncreased amount of new bone formation[129]
Hydroxyapatite –montmorillonite In vitroDecreased biodegradation, increased biomineralization and mechanical properties[130]
Nano SiO2 In vitroNo cytotoxicity, decreased water retention, increased protein adsorption, biomineralization, biodegradation, and mechanical properties[131]
β-tricalcium phosphate In vitroNo cytotoxicity, increased water retention, biomineralization, osteogenesis, and mechanical properties[132]
In vitroNo cytotoxicity, increased biomineralization and mechanical properties, decreased biodegradation[133]
Hydroxyapatite In vitroNo cytotoxicity, increased osteogenesis and mechanical properties[134]
Fucoidanβ-tricalcium phosphate In vitroNo cytotoxicity, increased protein adsorption, biomineralization, osteogenesis, and mechanical properties[135]
poly(propylene carbonate) In vitroNo cytotoxicity, increased mechanical properties[136]
xybutyrate-co3-hydroxyvalerate (PHBV)
Hydroxyapatite In vitroNo cytotoxicity, increased biomineralization, osteogenesis, and mechanical properties[137]
Polyvinyl pyrrolidoneBioactive glass In vitroNo cytotoxicity, decreased biodegradation[138]
Polypyrrole-alginate In vitroNo cytotoxicity, increased biomineralization, decreased water retention, protein adsorption, and biodegradation[139]
alcohol or collagen
Bioactive glass In vitroNo cytotoxicity, increased biomineralization and mechanical properties, decreased water retention and biodegradation[140]
In vitroNo cytotoxicity, increased water retention, osteogenesis, and mechanical properties[141]
In vivoGood cartilage healing in rabbit
Polylactide + AlginateVEGFIn vitroGood VEGF release rate, enhanced neovascularization in bone healing and maintenance of bioactivity[142]
In vivo
Table 2. Chitosan-based scaffold in dental-pulp regeneration.
Table 2. Chitosan-based scaffold in dental-pulp regeneration.
Chitosan-Based Biocomposite Scaffolds
Polymers and/or BiomaterialsBioactive MoleculeModelObservationsReference
β-tricalcium phosphate In vitro
  • Upregulated expressions of alkaline phosphatase (ALP) and osteopontin (OPN)
CollagenBMP-7In vivo
  • Release of BMP-7 gene
  • Dental pulp stem cells (DPSC) differentiation into odontoblast-like cells in vitro and in vivo
Calcium-aluminate1α,25-dihydroxyvitamin D3 (1α,25VD)In vitro
  • Increased odontoblastic phenotype expression
  • Cell migration
Fibrin In vitro
  • Potent antibacterial effect
  • Similar dental pulp-mesenchymal stem cells (DP-MSC) viability, fibroblast-like morphology, proliferation rate
  • Type I/III collagen production capacity.
Silver-doped bioactive glass In vitro
  • The proliferation of dental pulp cells (DPC) is not affected
  • Decrease of inflammation
  • Odontogenic differentiation of DPCs
  • Inhibition of Streptococcus mutans and Lactobacillus casei growth
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