Functionalization Strategies of Chitosan-Based Scaffolds with Growth Factors for Bone Regeneration: A Systematic Review
Abstract
1. Introduction
2. Results
2.1. Study Selection
2.2. General Characteristics of the Included Studies
2.3. Main Study Outcomes
2.3.1. Histological and Imaging Outcomes
2.3.2. Enzymatic Activity (ALP, OCN, PDGF, ELISA)
2.3.3. Gene Expression of Osteogenic Markers
2.3.4. Mineralization and Calcium Deposition
2.3.5. Cell Proliferation and Morphology
2.3.6. Angiogenesis and VEGF Expression
2.3.7. Release Kinetics and Delivery Strategies
2.4. Quality Assessment
2.5. Risk of Bias in Included Studies
2.6. Reporting Biases
3. Discussion
4. Materials and Methods
4.1. Focused Question:
4.2. Protocol
4.3. Eligibility Criteria
- Studies examining the effect of adding growth factors to chitosan scaffolds on bone regeneration.
- In vitro studies
- In vivo studies
- Studies with a control group;
- Studies in English;
- Non-randomized controlled clinical trials (NRS); and
- Randomized controlled clinical trials (RCT).
- Non-English papers;
- Clinical reports;
- Opinions;
- Editorial papers;
- Review articles;
- No full-text accessible; or
- Duplicated publications.
4.4. Information Sources, Search Strategy, and Study Selection
4.5. Data Collection Process and, Data Items
4.6. Risk of Bias and Quality Assessment
4.7. Quality Assessment
- Is it clear in the study what is the “cause” and what is the “effect”?
- Were the participants included in any similar comparisons?
- Were the participants included in any comparisons receiving similar treatment/care, other than the exposure or intervention of interest?
- Was there a control group?
- Were there multiple measurements of the outcome both before and after the intervention/exposure?
- Was a follow up completed, and if not, were the differences between groups in terms of their follow up adequately described and analyzed?
- Were the outcomes of the participants included in any comparisons measured in the same way?
- Were the outcomes measured in a reliable way?
- Was an appropriate statistical analysis used?
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Conflicts of Interest
References
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Bioactive Factor | Main Biological Role | Functionalization Strategy (Chemical/Physical) | References |
---|---|---|---|
BMP-2 | Potent osteoinductive factor; promotes differentiation of mesenchymal stem cells into osteoblasts | Grafting (P24 peptide), covalent coupling, microsphere encapsulation, nanoparticle incorporation, heparinization | [51,52,54,62,63,76,77,78,79,80] |
BMP-6 | Strong osteogenic factor; stimulates osteoblast activity and matrix mineralization | Encapsulation in alginate microspheres or PHBV submicron particles | [55,81] |
IGF-1 | Stimulates proliferation and differentiation of osteoblasts; enhances collagen synthesis | Physical adsorption, soaking into CS scaffolds | [77] |
VEGF | Stimulates angiogenesis and vascularization within regenerating tissue | Encapsulation in alginate or PLGA microspheres, composite reinforcement (e.g., Aloe vera, HA-CS) | [54,82,83,84] |
bFGF (FGF-2) | Enhances proliferation of mesenchymal stem cells and angiogenesis; delays differentiation | Embedding in CS–HA scaffolds, release control via HA interactions | [85] |
PDGF | Promotes proliferation and angiogenesis; recruits progenitor cells | Combined encapsulation (alginate microspheres, brushite–CS scaffolds) | [81,84] |
Periostin | Extracellular matrix protein; supports adhesion, migration, and mechanical stress adaptation | Physical adsorption on genipin–crosslinked chitosan | [56] |
Study | Aim of the Study | Material and Methods | Results | Conclusions |
---|---|---|---|---|
Wang et al. [54] | To determine whether an integrated, injectable hydrogel platform delivering BMP-2, VEGF, and adipose-derived mesenchymal cells (ADSCs) supports the regeneration of vascularized bone tissue in rabbit mandibular defects. | Chitosan hydrogel with PLGA/nHA microspheres containing BMP-2 and VEGF was prepared and implanted together with ADSCs into a critical bone defect in the mandible of rabbits; bone lesion formation was assessed by computed tomography and macro- and histological observations for 12 weeks | Hydrogel with BMP-2 and VEGF + ADSC significantly accelerated bone formation, shortened healing time, and improved callus remodeling compared to the control and VEGF-alone groups, with full defect filling after 12 weeks | Co-delivery of VEGF and BMP-2 along with ADSCs through chitosan hydrogel effectively promotes simultaneous osteogenesis and angiogenesis, making this strategy very promising for regenerative bone engineering. |
Yun et al. [62] | We investigated whether heparin-coated chitosan scaffolds capable of sustained release of BMP-2 improved osteoblast function compared to traditional chitosan or BMP-2/chitosan. | In vitro, the osteoblast cell line MG-63 cultured on three types of scaffolds (chitosan, BMP-2/chitosan, and BMP-2/heparin/chitosan) was used. Proliferation, ALP activity, calcium deposition, and expression of osteogenic genes were assessed. | BMP-2/heparin/chitosan scaffolds provided a more continuous BMP-2 efflux, which resulted in a significant increase in ALP activity and calcium deposition in osteoblast cells compared to the other groups | The heparin coating of the chitosan scaffold enables sustained release of BMP-2, which positively influences osteoblast function—a promising strategy for bone regeneration. |
Sanjaya et al. [76] | To check whether the use of a combination of rhBMP-2 and a chitosan scaffold accelerates the regeneration of calvarial defects in Wistar rats. | Twenty-four male Wistar rats were used, randomly assigned to 4 groups (control, rhBMP-2 only, chitosan scaffold only, rhBMP-2 + chitosan combination), and healing was assessed after 4 weeks by measuring osteocalcin expression, osteoblast number, fibrous bone area, and PDGF level. | The combination of rhBMP-2 and chitosan scaffold significantly increased osteocalcin expression, osteoblast number, fibrous bone area, and PDGF compared to single interventions and the control group | The combination of rhBMP-2 with chitosan scaffold effectively accelerates bone regeneration in skull defects in rats, exceeding the effects of rhBMP-2 or chitosan alone |
Tigli et al. [85] | To assess whether chitosan scaffolds containing dexamethasone or bFGF with hydroxyapatite can provide controlled, sustained drug release for tissue engineering purposes. | Pure chitosan or chitosan-HA scaffolds were prepared by incorporating different doses of dexamethasone (300–900 ng) or bFGF (50–100 ng) per dry sample (3 mg) and analyzing the release in DPBS buffer in vitro for ≤5 days using UV-VIS spectrophotometry. | The scaffolds demonstrated continuous release of dexamethasone for approximately 5 days, while bFGF was completely released within 10–20 h. The addition of hydroxyapatite allowed for prolonged bFGF release to up to 7 days due to electrostatic interactions with HA. | The chitosan-HA design enables controlled and sustained release of both dexamethasone and bFGF, making these materials a promising solution for applications in bone and periodontal tissue regeneration. |
Liu et al. [83] | Development and testing of a novel composite scaffold of hydroxyapatite, sodium alginate and chitosan capable of simultaneously releasing vancomycin and VEGF to accelerate bone regeneration and prevent infections. | The scaffolds were encapsulated within a VEGF microsphere with a vancomycin-containing layer and their physical properties, drug release rate, in vitro bactericidal effect against S. aureus, and osteogenic potential of MSCs were tested. | The HA6(SA/CS)4@VAN/VEGF scaffold exhibited optimal porosity, controlled degradation, dual drug release within the designed time frame, potent antibacterial activity against S. aureus, good biocompatibility, and support for MSC proliferation and osteogenesis | The HA-SA-CS composite scaffold with embedded VEGF and vancomycin shows promising potential as a bone tissue engineering material, combining mechanical, antibacterial, and pro-osteogenic properties. |
Barakzai et al. [56] | To investigate whether chitosan implants containing periostin accelerate bone regeneration in the absence of mechanical loading. | The study involved 36 rats divided into three groups (control, chitosan, and chitosan with periostin). Scaffolds were implanted into femoral defects, and regeneration was assessed histologically and morphometrically after 4 and 8 weeks. | The periostin group showed significantly greater bone regeneration, better tissue organization, and increased expression of osteogenesis markers compared to the other groups. | The addition of periostin to a chitosan scaffold effectively promotes bone healing under non-weight-bearing conditions, suggesting the clinical potential of this combination in the treatment of bone defects. |
Chen [51] | Investigate whether the CS-P24/HA scaffold, which delivers the P24 peptide from BMP2, can be used in bone regeneration procedures. | P24 and BMP were incorporated into the chitosan structure. HA was then added to form the CS-5%P24/HA or CS-10%P24/HA scaffold. CS-HA was used as a control. In vitro, the release of P24 from the samples into a PBS solution was assessed chromatographically over 90 days. Quantitative assessment by HPLC. Rat bone marrow stem cells were seeded onto the CS-5%P24/HA and CS-10%P24/HA scaffolds. Cell proliferation, ALP activity, Ca deposition and osteogenic mRNA levels were assessed. In vivo: Three groups of rats were implanted with CS/HA, CS-5%P24/HA and CS-10%P24/HA. After sacrifice, scaffold samples were analyzed by microtomography. Assessment: BMC, BMD, TMD, histology and immunohistochemistry (expression of: Ocn, Nestin and CD31. In vivo no. The following were implanted into the skull defects of rats: CS/HA, CS-5%P24/HA and CS-10%P24/HA. After sacrifice, the study was conducted as in No. 1. | In vitro: linear and stable release of P24 from both test samples over 90 days. In vivo: stem cell count, alkaline phosphatase (ALP) activity, cell proliferation, mRNA levels, calcium (Ca) levels, quantity and quality of regenerated bone were all higher with CSP24/HA than with CS/HA. | P24-containing chitosan scaffolds promote faster and better bone regeneration. |
Soriente [52] | The creation of a bioactive chitosan scaffold that enhances osteogenesis. | Two types of bioactive chitosan scaffold were created: 1) with BMP (organic), and 2) with HA nuclei (inorganic). The scaffold structure was assessed using SEM and MicroCT. In vitro: Peptide release from the BMP scaffolds was analyzed by chromatography. Cell proliferation on the scaffolds was evaluated using the Alamar blue assay. ALP activity was assessed on hMSC scaffolds. Quantitative OCN measurements from the substrate were used as a marker of osteogenesis. | Higher ALP and OCN values were observed for the tested scaffolds containing BMP and biomineralized HA. The higher the concentration of BMP2, the better the cell morphology, i.e., the structure of osteoblasts. At lower concentrations, the morphology resembles that of fibroblasts. Faster cell proliferation was observed in the presence of BMP2. | Adding BMP2 and HA to chitosan scaffolds may encourage bone regeneration. |
Soran [55] | The effect of adding alginate microspheres containing BMP-6 to chitosan gels on periodontal tissue regeneration. | Chitosan scaffolds containing alginate microspheres were prepared (1. empty; 2. enriched with BMP-6). In vitro: BMP-6 release measurement was performed using a fluorescence spectrophotometer. rBM-MSC cells were injected into: 1. CS, 2. CS + BMP-6, and 3. CS + BMP-6 + alginate. ALP and mineralization were analyzed using van Kossy, and cell morphology was assessed using SEM. | The most intense stem cell proliferation was observed in the BMP-6 plus microspheres group. The ALP level was highest in the BMP-6 plus microspheres group, significantly higher than in the free BMP-6 group. Calcification was also highest in this group. The BMP-6 plus microspheres group also exhibited the most complex stem cell morphology. | Chitosan scaffolds containing alginate microspheres and BMP-6 demonstrated the most effective delivery of factors that promote stem cell development. |
Nandi [77] | How does the addition of IGF-1 and BMP-2 to a chitosan scaffold affect bone healing | Chitosan scaffolds were impregnated with IGF-1 and BMP-1. In vivo: rabbit tibiae with defects were filled with the following: A. no treatment, B. chitosan + IGF-1, or C. chitosan + BMP-1. Histopathological analysis was performed after 30 and 90 days following implant placement. Bone sections were analyzed under an Orthoplan microscope. The sections were also examined using SEM. | Compared to group A, there was greater osteoblast activity, a greater number of cells, and increased proliferation of bone tissue and vessels in groups B and C. Bone formation density was also significantly higher in groups B and C. Fluorescence in the microscopic image was much more intense in groups B and C. | The addition of IGF-1 and BMP-1 to chitosan significantly accelerates the regeneration and reconstruction of bone tissue. |
Sularsih [82] | Analysis of the pores-size of chitosan-Aloe vera scaffold on alveolar bone healing and VEGF expressions after tooth removal (study was conducted on Cavia cobaya). | The study was conducted on 36 males of Cavia cobaya, divided into 6 groups (n = 6): negative control (without scaffolds), positive control (chitosan scaffold), treatment groups (chitosan–aloe vera scaffold). Data analyzed: Woven alveolar bone areas and VEGF expressions (by histopathological examination, ANOVA, LSD) Scaffold pore size (by SEM) | The largest woven alveolar bone and the highest expression on VEGF was observed in the treatment groups after 7 and 14 days (statistically significant difference between control and treatment group). Open pore interconnectivity was observed in chitosan- Aloe vera scaffolds. | Usage of chitosan–aloe vera scaffold (by its pore characteristics) increased VEGF expressions and woven alveolar bone areas. |
Yun [63] | Development of novel bone-grafting scaffolds (BMP-2-immobilizingheparinized-chitosan scaffolds) And assessment of its osteogenic differentiation activity. | Chitosan scaffolds were functionalized with heparin and BMP-2 to enhance osteogenic potential. Measured factors:
| The results showed that BMP2-immobilizing heparinized-chitosan (BMP-2/Hep-chitosan) scaffolds significantly enhanced ALP activity and calcium deposition of the osteoblast cells when compared with chitosan scaffolds only. Also, mRNA expressions of osteocalcin and osteopontin of osteoblast cells cultured on BMP-2 (100 ng)/Hep-chitosan scaffolds were increased compared to chitosan scaffolds. | BMP-2/Hep-chitosan systems can increase osteoblast activity, therefore such systems are a platform to develop next generation transplant materials. |
Guzman [78] | Evaluation of osteoinductive capability of the immobilized bone morphogenetic protein adsorbed to chitosan scaffold after manufacturing process. | Materials: chitosan (CHI), hydroxyapatite (HAp), urease, urea, and rhBMP-2. Hydrogel Preparation: CHI hydrogels were mixed with urea, urease, and optionally rhBMP-2 or CPS, then gelled at 37 °C for 24 h. ISISA Processing: Hydrogels were frozen, freeze-dried, and cryo-fractured into 2.5 mm scaffolds. In Vitro Assays:
| The results showed that scaffolds containing both rhBMP-2 and CPS-CHI enhanced bone regeneration, with rhBMP2-CPS-CHI scaffolds inducing significantly more new bone formation compared to the other scaffolds. While rhBMP2-CHI alone did not lead to increased bone growth, the combination of CPS with rhBMP-2 promoted a more effective release of the growth factor, resulting in superior osteoinductive effects. Histological and microCT analyses confirmed the presence of trabecular bone and chondral zones in defects treated with rhBMP2-CPS-CHI. | rhBMP2 was released in a controlled way from scaffold and retained its osteoinductive character after release. The multicomponent scaffold showed better regenerative abilities than the scaffolds containing only one of the components (CPS or rhBMP2 separately). |
De la Riva [84] | Evaluation the effectiveness of a brushite- chitosan scaffold loaded with PDGF or a combination of PDGF and VEGF in supporting bone regeneration. | Brushite–chitosan scaffolds were developed, with PDGF combined in the cement liquid phase and VEGF encapsulated in alginate microspheres embedded in chitosan sponges. In vitro and in vivo release kinetics were examined using radiolabeled growth factors (125I). Implants were inserted into rabbit femur with bone defect and analyzed for growth factor release, distribution and bone regeneration. | Both in the in vivo and in vitro study PDGF was released faster than VEGF. Both factors stayed localized. Bone regeneration was enhanced in scaffolds containing PDGF and even more in those containing both—PDGF and VEGF. | PDGF supports early stages of healing, while VEGF delivery aids in later bone formation and vascularization. Bone regeneration after dual delivery of GF (PDGF + VEGF) was higher than single GF application. The brushite-chitosan scaffold allows localized, time- controlled release, supporting tissue regeneration. |
Demirtaş [81] | This study aimed was to evaluate the positive effects of combined delivery of two growth factors—PDGF-BB (a mitogenic factor) and BMP-6 (an osteogenic factor)—loaded within a chitosan-based scaffold, on growth and maturation of pre-osteoblastic MC3T3-E1 cells. | PDGF-BB was loaded into gelatin microparticles and BMP-6 into PHBV submicron particles. These were then incorporated into 3D porous chitosan scaffolds prepared by freeze-drying. Different degradation rates led to faster PDGF-BB and slower BMP-6 release. MC3T3-E1 cells were seeded onto the scaffolds and analyzed over 21 days for viability, proliferation (MTT), morphology (SEM), and gene expression (RT-PCR). | Chitosan scaffolds loaded with both PDGF-BB and BMP-6 promoted greater cell proliferation and significantly enhanced expression of osteogenic markers (RunX2, Col1, OPN, OCN) compared to single-factor scaffolds. PDGF-BB was released rapidly, enhancing early cell proliferation, while BMP-6 had a sustained release supporting differentiation. | Chitosan serves as a biocompatible scaffold but needs incorporation of growth factors to promote osteogenesis effectively. The dual- release system using PDGF-BB and BMP-6 in chitosan scaffolds showed more effective than single-factor systems. This approach supports physiological mechanism of tissue repair. |
Xie [80] | The aim of the study was to develop a biomimetic scaffold based on chitosan and graphene oxide, mineralized with octacalcium phosphate (OCP), and functionalized with bone morphogenetic protein-2 (BMP-2) and silver nanoparticles (Ag-NPs). The research focused on evaluating whether incorporating BMP-2 into the scaffold would improve its osteoinductive capacity and promoting bone tissue regeneration. | Porous scaffolds composed of GO and chitosan were fabricated and then mineralized using a supersaturated calcium phosphate solution to deposit OCP. BMP-2 was encapsulated in BSA nanoparticles stabilized with chitosan and immobilized on the scaffold via adsorption. Characterization included structural analysis, BMP-2 release studies, in vitro tests with bone marrow stromal cells (BMSCs), and in vivo implantation in rat cranial defect models. | Scaffolds with BMP-2-loaded nanoparticles exhibited sustained release of the growth factor, significantly enhancing BMSC proliferation and osteogenic differentiation compared to controls. In vivo, these scaffolds promoted more extensive new bone formation. The delivery of BMP-2 from nanoparticles was more effective than direct surface adsorption. | Incorporating BMP-2 into OCP-GO/CS scaffolds via nanoparticle encapsulation notably improved their osteoinductive properties. The controlled release of BMP-2 created a favorable microenvironment for bone regeneration. |
Qiu [79] | The study aimed to develop a biomaterial-based scaffold that enables controlled and sustained release of—BMP-2, enhancing its biological effectiveness in bone regeneration. | Three types of scaffolds were prepared: one without BMP-2 (SCH), one with BMP-2 simply adsorbed on the surface (SCH-D), and one where BMP-2 was preloaded into mHANPs and then embedded in the scaffold (SCH-L). All were assessed through in vitro tests (release kinetics, ALP activity, gene expression) and in vivo in rat skull defects. | SCH-L showed a significantly slower initial BMP-2 release and higher retention of its bioactivity than SCH-D. In cell studies, SCH-L led to significantly higher ALP activity and gene expression (RunX2, Col I, ALP, OC). In vivo, SCH-L generated more uniform and extensive new bone formation. | Preloading BMP-2 into mHANPs resulted in a more effective and bioactive release profile. The SCH-L scaffold promoted stronger in vitro and in vivo bone regeneration. |
Authors | Study Model | Scaffold Composition | Bioactive Agent and Functionalization Strategy | Bone Regeneration Assessment Methods | Observed Effects on Bone Regeneration |
---|---|---|---|---|---|
Wang et al. [54] | In vivo, rabbit | Chitosan (CS) + nanohydroxyapatite (nHA) + PLGA microspheres | BMP-2/VEGF-loaded PLGA microspheres | 3D-CT, histology (Masson’s stain), ALP, Ca deposition, in vitro bioactivity assessment |
|
Yun et al. [62] | In vitro—MG-63 cell line (osteoblasts) | CS + heparin (coating) | Chemically immobilized BMP-2 on a heparinized scaffold surface | ALP, Ca deposition, gene expression (OCN, OPN), cell proliferation (CCK-8) |
|
Sanjaya et al. [76] | In vivo, rat | Chitosan gel 3% in HPMC 5% | Co-administration/co-incorporation rhBMP-2 | Histology (H&E)—osteoblast count and % woven bone tissue; ELISA: osteocalcin (OCN) and PDGF | significantly higher values of OCN, PDGF, % woven bone and osteoblast number compared to the control group |
Tigli et al. [85] | In vitro | CS + HA | bFGF embedding/solvent sorption method | Dexamethasone and bFGF release study, fluorescence, UV, release kinetics | without HA—bFGF release in 10–20 h; with HA—extended to 7 days due to electrostatic interactions with HA |
Liu et al. [83] | In vitro, rat | HA/sodium alginate (SA)/CS | Multilayer microspheres with VEGF inside | Adhesion (confocal), ALP (staining + semi-quantitative), mineralization (Alizarin Red), gene expression (qPCR: ALP, BMP2, OPN, RunX2); VEGF/VAN release; antibacterial tests (OD and inhibition zones) | greater cell adhesion, higher ALP activity, stronger mineralization and higher ALP/BMP2/OPN/RunX2 expression compared to the control group |
Barakzai et al. [56] | In vivo, rat | Chitosan (2% w/v) cross-linked with genipin | Periostin Surface deposition/adsorption | Micro-CT, histology (H&E, toluidine, Masson’s trichrome), ALP activity | Higher osteocyte count, higher bone volume and collagen fiber percentage compared to the control group, good cell viability and degradation in vitro. Periostin accelerated regeneration under unloaded conditions. |
Chen [51] | In vitro, rat BMSC In vivo, rat | CS-HA | P24 from BMP-2, chemical grafting on chitosan scaffolds |
| Scaffolds with P24 comparing to control group (no P24):
|
Soriente [52] | In vitro, human stem cell line | -CS + polyethylene gly- col diacrylate (20%) -CS + polyethylene gly- col diacrylate (40%) | BMP-2, covalent coupling on scaffold surface |
| Scaffold with BMP-2 comparing to biomineralized scaffolds:
|
Soran [55] | In vitro, rat stem cells | -CS + alginate microspheres | BMP-6; BMP-6 in alginate microspheres mixed with CS |
| Group with alginate microspheres:
|
Nandi [77] | In vivo, rabbit | Chitosan-based scaffold | IGF-1; BMP-2; soaking the scaffolding |
| Groups with IGF-1 and BMP-2:
|
Sularsih [82] | In vivo, 36 male Cavia cobaya (guinea pigs), aged 3–3.5 months | chitosan–aloe vera scaffold | VEGF | SEM, HE and MICONOS, Antibody monoclonal VEGF | The highest VEGF expression and the largest woven alveolar bone were found in the treatment groups with chitosan–aloe vera scaffold. |
Yun [63] | In vitro, Human osteosarcoma MG-63 cells | BMP-2/Hep-Chitosan scaffolds | BMP-2- scaffolds incubated in MES buffer containing heparin and EDC for 24 h at 4 °C, immobilization of BMP-2 for 4 h. Sterilization in 70% ethanol and washing with PBS | SEM, ELISA, ALP, Ca deposition, RT-PCR | The expression levels of osteocalcin and osteopontin were upregulated in the BMP-2 (100 ng)/Hep-chitosan group Cell proliferation higher in BMP-2 (100 ng)/chitosan scaffold |
Guzman [78] | In vitro: C2C12 cell line In vivo: 20 New Zealand male rabbits of ca. 3 kg in weight | CHI, CPS-CHI, rhBMP2-CHI, and rhBMP2-CPS-CHI scaffolds | CPS, BMP-2—Urease-assisted preparation of chitosan hydrogels, ISISA process | SEM, Invitrogen, ALP, ELISA, Micro-CT, Histology and histomorphometry | Cells proliferation and osteoinduction higher in scaffolds with both CPS and rhBMP-2. |
De la Riva [84] | In vivo New Zealand rabbits, male Femur defects In vitro-release kinetics of GF | Brushite–chitosan scaffold | PDGF—dissolved in liquid phase of brushite, VEGF—encapsulated in alginate microspheres inside chitosan sponges, | Bone regeneration was assessed using in vivo methods:
| Osteogenesis: PDGF increased new bone formation and osteoblastic activity VEGF enhanced later-stage bone formation PDGF + VEGF combination led to the greatest bone area, larger trabeculae, and more osteoblast layers. Mineralization: Greater mineralized bone and osteoid matrix were observed, especially in the dual-factor group (VP). Cellular activity: Mitotic osteoprogenitor cells and multilayered osteoblasts were observed in the VEGF + PDGF group. Angiogenesis: Not directly measured. |
Demirtaş [81] | In vitro Cell culture: MC3T3-E1 mouse pre-osteoblast cell line | Chitosan-based scaffold |
| RT-PCR for gene expression of osteogenic markers: RunX2, Col1(Collagen type I), OPN (Osteopontin), OCN (Osteocalcin). SEM imaging MTT assay Calcium deposition observed via SEM/EDAX. | Osteogenesis: higher expression of RunX2, Col1, OPN, and OCN, especially with dual factor delivery, Cell proliferation: markedly increased on PDGF-BB-containing scaffolds, Mineralization: calcium deposits detected after 7 days in scaffolds with both growth factors, Cell attachment and morphology: improved spreading and cell–cell interaction observed on dual-loaded scaffolds. Angiogenesis: Not directly measured in this study. |
Xie [80] | In vitro and in vivo BMSCs—bone marrow stromal cells Sprague-Dawley rats | OCP-GO/CS scaffolds (octacalcium phosphate mineralized graphene oxide/chitosan scaffolds) | BMP-2-encapsulated BSA (bovine serum albumin) nanoparticles (CBB-NPs) |
| BMP-2 had a significant effect on promoting osteogenic differentiation, as evidenced by elevated ALP activity. It also enhanced BMSC proliferation and supported new bone formation in vivo. The study did not directly evaluate effects on angiogenesis or calcium deposition separately. |
Qiu [79] | In vitro—BMSCs—bone marrow mesenchymal stem cells, rats In vivo—SD rats, calvarial defect model—osteogenic effect | silk fibroin (SF) and chitosan (CS) | BMP-2 was immobilized using mesoporous hydroxyapatite nanoparticles (mHANPs). |
| BMP-2 significantly enhanced osteogenic differentiation, matrix mineralization, and cell attachment and proliferation in vitro. In vivo, BMP-2 promoted new bone formation within rat cranial defects. Angiogenesis was not evaluated in this study. |
Authors | 1. Is It Clear in the Study What Is the ‘Cause’ and What Is the ‘Effect’? | 2. Were the Participants Included in Any Comparisons Similar? | 3. Were the Participants Included in Any Comparisons Receiving Similar Treatment/Care, Other than the Exposure or Intervention of Interest? | 4. Was There a Control Group? | 5. Were There Multiple Measurements of the Outcome Both Pre and Post the Intervention/Exposure? | 6. Was Follow Up Complete and If Not, Were Differences Between Groups in Terms of Their Follow Up Adequately Described and Analyzed? | 7. Were the Outcomes of Participants Included in Any Comparisons Measured in the Same Way? | 8. Were Outcomes Measured in a Reliable Way? | 9. Was Appropriate Statistical Analysis Used? |
---|---|---|---|---|---|---|---|---|---|
Wang et al. [54] | yes | yes | yes | yes | yes | yes | yes | yes | yes |
Yun et al. [62] | yes | yes | yes | yes | yes | yes | yes | yes | yes |
Sanjaya et al. [76] | yes | yes | yes | yes | no | yes | yes | yes | yes |
Tigli et al. [85] | yes | yes | yes | yes | yes | yes | yes | yes | yes |
Liu et al. [83] | yes | yes | yes | yes | yes | yes | yes | yes | yes |
Barakzai et al. [56] | yes | yes | yes | yes | yes | yes | yes | yes | yes |
Chen [51] | yes | yes | yes | yes | yes | yes | yes | yes | yes |
Soriente [52] | yes | yes | yes | yes | yes | yes | yes | yes | yes |
Soran [55] | yes | yes | yes | yes | yes | yes | yes | yes | yes |
Nandi [77] | yes | yes | yes | yes | yes | yes | yes | yes | yes |
Sularsih [82] | yes | yes | yes | yes | yes | yes | yes | yes | yes |
Yun [63] | yes | yes | yes | yes | yes | yes | yes | yes | yes |
Guzman [78] | yes | yes | yes | yes | yes | yes | yes | yes | yes |
De la Riva [84] | yes | yes | yes | no | yes | yes | yes | yes | yes |
Demirtaş [81] | yes | yes | yes | yes | yes | yes | yes | yes | yes |
Xie [80] | yes | yes | yes | yes | yes | yes | yes | yes | yes |
Qiu [79] | yes | yes | yes | no | yes | yes | yes | yes | yes |
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Kiryk, J.; Michalak, M.; Majchrzak, Z.; Laszczyńska, M.; Kiryk, S.; Szotek, S.; Gerber, H.; Nawrot-Hadzik, I.; Matys, J.; Dobrzyński, M. Functionalization Strategies of Chitosan-Based Scaffolds with Growth Factors for Bone Regeneration: A Systematic Review. Mar. Drugs 2025, 23, 396. https://doi.org/10.3390/md23100396
Kiryk J, Michalak M, Majchrzak Z, Laszczyńska M, Kiryk S, Szotek S, Gerber H, Nawrot-Hadzik I, Matys J, Dobrzyński M. Functionalization Strategies of Chitosan-Based Scaffolds with Growth Factors for Bone Regeneration: A Systematic Review. Marine Drugs. 2025; 23(10):396. https://doi.org/10.3390/md23100396
Chicago/Turabian StyleKiryk, Jan, Mateusz Michalak, Zuzanna Majchrzak, Marzena Laszczyńska, Sylwia Kiryk, Sylwia Szotek, Hanna Gerber, Izabela Nawrot-Hadzik, Jacek Matys, and Maciej Dobrzyński. 2025. "Functionalization Strategies of Chitosan-Based Scaffolds with Growth Factors for Bone Regeneration: A Systematic Review" Marine Drugs 23, no. 10: 396. https://doi.org/10.3390/md23100396
APA StyleKiryk, J., Michalak, M., Majchrzak, Z., Laszczyńska, M., Kiryk, S., Szotek, S., Gerber, H., Nawrot-Hadzik, I., Matys, J., & Dobrzyński, M. (2025). Functionalization Strategies of Chitosan-Based Scaffolds with Growth Factors for Bone Regeneration: A Systematic Review. Marine Drugs, 23(10), 396. https://doi.org/10.3390/md23100396