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10.3390/cells15030215
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Article

Cell Supported Single Membrane Technique for the Treatment of Large Bone Defects: Depletion of CD8+ Cells Enhances Bone Healing Mechanisms During the Early Bone Healing Phase

by
Marissa Penna-Martinez
*,
Lia Klausner
,
Andreas Kammerer
,
Minhong Wang
,
Alexander Schaible
,
René Danilo Verboket
,
Christoph Nau
,
Ingo Marzi
and
Dirk Henrich
Department of Trauma Surgery and Orthopedics, University Hospital, Goethe University Frankfurt, 60590 Frankfurt am Main, Germany
*
Author to whom correspondence should be addressed.
Cells 2026, 15(3), 215; https://doi.org/10.3390/cells15030215
Submission received: 2 December 2025 / Revised: 15 January 2026 / Accepted: 20 January 2026 / Published: 23 January 2026
(This article belongs to the Special Issue New Advances in Tissue Engineering and Regeneration)
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Abstract

Introduction: The one-step membrane technique, derived from the Masquelet induced membrane technique, uses human acellular dermal matrix (hADM) that is wrapped around the bone defect to bypass membrane induction, reducing treatment time. Pre-colonization of hADM with bone marrow cells (BMC), particularly after CD8+ T cell depletion, enhances bone regeneration. This study examined how CD8+ T cell depletion alters the proteins accumulated in the hADM during early healing. Materials and Methods: Eighteen male Sprague-Dawley rats received 5 mm femoral defects filled with autologous bone chips and wrapped with hADM, hADM + BMC, or hADM + BMC-CD8. hADMs were recovered on days 3 and 7 (n = 3/group/timepoint), incubated ex vivo, and conditioned medium analyzed with a proteome profiler detecting 79 proteins. Results: The protein content of the hADM evolved dynamically. At day three, 41 proteins were detected, rising to 47 by day seven, with RGM-A, osteoprotegerin, LIF, IL-6, CCL20, and CCL17 emerging late, consistent with increased regenerative activity. CD8+ T cell depletion suppressed early inflammatory and pro-osteogenic mediators (e.g., CCL2, IGF-I, IL-1RA) while upregulating LIX. By day seven, regenerative mediators (CCL20, GDF-15, RGM-A) were enriched, whereas inflammatory factors (CCL21, IL-1a, WISP-1) declined. MMP-9, Galectin-1, and GDF-15 increased exclusively in the CD8-depleted group. Conclusions: The hADM protein content transitions from pro-inflammatory to pro-regenerative within one week after surgery. CD8+ T cell depletion accelerates this shift, highlighting hADM as a dynamic scaffold that contributes to the immune–regenerative crosstalk in bone healing.
Keywords: bone marrow mononuclear cells; BMC; CD8 lymphocytes; monocytes; mesenchymal stem cells; large bone defect; early bone healing; acellular dermis bone marrow mononuclear cells; BMC; CD8 lymphocytes; monocytes; mesenchymal stem cells; large bone defect; early bone healing; acellular dermis

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MDPI and ACS Style

Penna-Martinez, M.; Klausner, L.; Kammerer, A.; Wang, M.; Schaible, A.; Verboket, R.D.; Nau, C.; Marzi, I.; Henrich, D. Cell Supported Single Membrane Technique for the Treatment of Large Bone Defects: Depletion of CD8+ Cells Enhances Bone Healing Mechanisms During the Early Bone Healing Phase. Cells 2026, 15, 215. https://doi.org/10.3390/cells15030215

AMA Style

Penna-Martinez M, Klausner L, Kammerer A, Wang M, Schaible A, Verboket RD, Nau C, Marzi I, Henrich D. Cell Supported Single Membrane Technique for the Treatment of Large Bone Defects: Depletion of CD8+ Cells Enhances Bone Healing Mechanisms During the Early Bone Healing Phase. Cells. 2026; 15(3):215. https://doi.org/10.3390/cells15030215

Chicago/Turabian Style

Penna-Martinez, Marissa, Lia Klausner, Andreas Kammerer, Minhong Wang, Alexander Schaible, René Danilo Verboket, Christoph Nau, Ingo Marzi, and Dirk Henrich. 2026. "Cell Supported Single Membrane Technique for the Treatment of Large Bone Defects: Depletion of CD8+ Cells Enhances Bone Healing Mechanisms During the Early Bone Healing Phase" Cells 15, no. 3: 215. https://doi.org/10.3390/cells15030215

APA Style

Penna-Martinez, M., Klausner, L., Kammerer, A., Wang, M., Schaible, A., Verboket, R. D., Nau, C., Marzi, I., & Henrich, D. (2026). Cell Supported Single Membrane Technique for the Treatment of Large Bone Defects: Depletion of CD8+ Cells Enhances Bone Healing Mechanisms During the Early Bone Healing Phase. Cells, 15(3), 215. https://doi.org/10.3390/cells15030215

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