Preparation and characterization of non-osteogenic compact bone scaffold for tendon tissue engineering
Year: 2016
Type: article
Abstract: Event Abstract Back to Event Preparation and characterization of non-osteogenic compact bone scaffold for tendon tissue engineering Quan Qing1, 2*, Yan-Jing Zhang1*, Jie-Liang Yang1*, Liang-Ju Ning1*, Ya-Jing Zhang1, Yi Zhang1*, Jing-Cong Luo1* and Ting-Wu Qin1 1 West China Hospital, Sichuan University, Regenerative Medicine Research Center, China 2 Sichuan College of Traditional Chinese Medcine, China Introduction: Demineralized bone matrix (DBM) possesses two critical characteristics of composition. First, collagen, sustains the structure and biomechanical strength of DBM and the same phenomenon is also presented in tendon and ligament. Second, non-collageonous proteins, undertake almost all biological functions of DBM, such as osteogenesis, the greatest advantage of DBM, which is relied on BMP-2. Researchers had studied the potential of DBM for tendon and ligament reconstruction[1],[2]. But the osteogenic ability of DBM which is the biggest barrier for soft tissue respair is not mentioned usually. DePaula[3] found that hydrogen peroxide (H2O2) could decrease the osteogenesis of DBM. Therefore, a hypothesis is proposed: on the basis of decellularized and demineralized compact bone matrix (DCDBM), whether a noval scaffold without osteogenic ability could be harvested after the treatment of H2O2. Comparing to DCDBM, the H2O2 treated bone matrix (HPTBM) should possesses the similar collagen structure and very significant decrease of osteogenesis. Materials and Methods: Bone matrix was harvested from limbs of newly born Simmental. A series of treatments, including decalcification, degreasing and decellularization, were applied to prepare DCDBM. A 12h soaking of 3% H2O2 was used for fabrication of HPTBM. The micro-structure and collagen structure of the two groups were observed with SEM, HE and Masson staining. Retained collagen content was detected with a hydroxyproline quantitative kit. Biomechanical properties were tested using a mechanical testing machine (Instron 8874). Co-culture with NIH 3T3 fibroblasts was used to evaluate the cytotoxicity. Immunohistochemitry (IHC) displayed the situation of reserved BMP-2. Bone marrow stromal cells of SD rat (rBMSCs) were seeded onto the surface of each sample from two groups and the expressions of osteopotin (OPN), osteocalcin (OCN), osteonectin (ON), were examined by RT-PCR in order to estimate the osteogenesis in vitro. rBMSCs grown in 6-well plates served as control. Results and Discussion: There was no significant difference of micro-structure and collagen structure between DCDBM and HPTBM. Collagen content was very close in both groups (Fig. 1A). The difference of UTS, strain at UTS, elastic modulus and stiffness of two groups was not significant (Fig. 1A). Both of two groups possessed the same cytotoxic grade, without significant cytotoxicity (Fig. 1C-H). IHC showed the higher BMP-2 in DCDBM (Fig. 2A&B). ALP staining revealed the higher ALP expression of rBMSCs seeded on DCDBM (Fig. 2C&D). The osteogenic gene expression analysis in vitro demonstrated that all the genes were regulated down for rBMSCs seeded on HPTBM (Fig. 2E). Due to the well preserved collagen, HPTBM got the similar biomechanical strength to DCDBM. With sufficient washing, the cytotoxicy of both scaffolds can be eliminated completely. BMP-2 may be washed away by H2O2, so that the signal path which is mediated by BMP-2 is blocked or blunted. This may directly lead to the down regulation of osteogenic gene expression of rBMSCs. Conclusion: HPTBM has a complete structure of collagen, good biomechanical properties, good cell compatibility and remarkable reduction of osteogenesis. This novel scaffold possesses the potential for tendon and ligament reconstruction. This work was supported by the grants from National High Technology Research and Development Program of China (2012AA020502) and National Natural Science Foundation of China (31370988).References:[1] Takeki Y, et al. J Biomech Sci Eng, 2007. 2:23-33.[2] Darryl A, et al. J Orthop Surg Res, 2013. 8:1-11.[3] DePaula C, et al. Cell Tissue Bank, 2005. 6:287-298. Keywords: Gene Expression, Tissue Engineering, mechanical property, acellullar matrix Conference: 10th World Biomaterials Congress, Montréal, Canada, 17 May - 22 May, 2016. Presentation Type: Poster Topic: Regenerative medicine: biomaterials for control of tissue induction Citation: Qing Q, Zhang Y, Yang J, Ning L, Zhang Y, Zhang Y, Luo J and Qin T (2016). Preparation and characterization of non-osteogenic compact bone scaffold for tendon tissue engineering. Front. Bioeng. Biotechnol. Conference Abstract: 10th World Biomaterials Congress. doi: 10.3389/conf.FBIOE.2016.01.01147 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 27 Mar 2016; Published Online: 30 Mar 2016. * Correspondence: Dr. Quan Qing, West China Hospital, Sichuan University, Regenerative Medicine Research Center, Chengdu, China, Email1 Dr. Yan-Jing Zhang, West China Hospital, Sichuan University, Regenerative Medicine Research Center, Chengdu, China, Email2 Dr. Jie-Liang Yang, West China Hospital, Sichuan University, Regenerative Medicine Research Center, Chengdu, China, Email3 Dr. Liang-Ju Ning, West China Hospital, Sichuan University, Regenerative Medicine Research Center, Chengdu, China, Email4 Dr. Yi Zhang, West China Hospital, Sichuan University, Regenerative Medicine Research Center, Chengdu, China, Email5 Dr. Jing-Cong Luo, West China Hospital, Sichuan University, Regenerative Medicine Research Center, Chengdu, China, Email6 Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Quan Qing Yan-Jing Zhang Jie-Liang Yang Liang-Ju Ning Ya-Jing Zhang Yi Zhang Jing-Cong Luo Ting-Wu Qin Google Quan Qing Yan-Jing Zhang Jie-Liang Yang Liang-Ju Ning Ya-Jing Zhang Yi Zhang Jing-Cong Luo Ting-Wu Qin Google Scholar Quan Qing Yan-Jing Zhang Jie-Liang Yang Liang-Ju Ning Ya-Jing Zhang Yi Zhang Jing-Cong Luo Ting-Wu Qin PubMed Quan Qing Yan-Jing Zhang Jie-Liang Yang Liang-Ju Ning Ya-Jing Zhang Yi Zhang Jing-Cong Luo Ting-Wu Qin Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.
Authors: Qing Quan, Zhang Yan-Jing, Yang Jie-Liang, Ning Liang-Ju, Zhang Ya-Jing, Zhang Yi, Luo Jing-Cong, Qin Ting-Wu
Language: English
Subfield: Biomedical Engineering
Field: Engineering
Domain: Physical Sciences
Open Access status: gold
Estimated APC paid: $3,801
OpenAlex ID: W2335819142