Abstract
3D porous hydroxyapatite (HA) has been reinforced by zirconia (ZrO2) coating and impregnation with a combination of platelet rich plasma (PRP) as a source of growth factors (GFs) and Heparin sulfate (HS) to sustain the release of GFs. Adipose mesenchymal stem cells (ADMSCs) were characterized by flow cytometry for CD (cluster of differentiation) 44, CD105, CD106, CD34 and CD144, along with checking the multipotency by differentiation into the adipocytes and osteoblasts. Then, they were cultured on the scaffold treated with and without osteogenic media on days 7, 14 and 21. Electron micrograph and PKH staining show that the ADMSCs have a fusiform phenotype in the absence of osteogenic induction. Cell viability assay shows a higher number of the viable cells on the PRP-containing scaffolds than PRP-free scaffolds on day 7. Colorimetric evaluation, quantitative RT-PCR and immunocytochemistry demonstrate that PRP and HS significantly elevate the alkaline phosphatase enzyme activity and also accelerate the production of both early and mid-osteogenic markers, including collagen I and osteopontin expression with and without osteogenic conditions. The PRP-HS also accelerates the expression of the late osteogenic marker, osteocalcin, in both mRNA and protein level expression with a peak on day 21. In conclusion, supplementation of HA/ZrO2 with PRP/HS has a synergistic impact on the ADMSCs, even in the absence of chemical induction. It seems that HA/ZrO2/PRP/HS scaffold provides a higher osteoconductive microenvironment for stem cell differentiation to osteoblasts.
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References
Afzal A (2014) Implantable zirconia bioceramics for bone repair and replacement: a chronological review. Mater Express 4:1–12. https://doi.org/10.1166/mex.2014.1148
Betsch M et al (2013) Bone marrow aspiration concentrate and platelet rich plasma for osteochondral repair in a porcine osteochondral defect model. PLoS ONE 8:e71602. https://doi.org/10.1371/journal.pone.0071602
Chen L, Lu X, Li S, Sun Q, Li W, Song D (2012) Sustained delivery of BMP-2 and platelet-rich plasma-released growth factors contributes to osteogenesis of human adipose-derived stem cells. Orthopedics 35:e1402–e1409. https://doi.org/10.3928/01477447-20120822-29
Chen D, Shen H, He Y, Chen Y, Wang Q, Lu J, Jiang Y (2015) Synergetic effects of hBMSCs and hPCs in osteogenic differentiation and their capacity in the repair of critical-sized femoral condyle defects. Mol Med Rep 11:1111–1119. https://doi.org/10.3892/mmr.2014.2883
Choi B-H, Im C-J, Huh J-Y, Suh J-J, Lee S-H (2004) Effect of platelet-rich plasma on bone regeneration in autogenous bone graft. Int J Oral Maxillofac Surg 33:56–59
Curran DJ, Fleming TJ, Towler MR, Hampshire S (2010) Mechanical properties of hydroxyapatite–zirconia compacts sintered by two different sintering methods. J Mater Sci Mater Med 21:1109–1120
Dombrowski C et al (2009) Heparan sulfate mediates the proliferation and differentiation of rat mesenchymal stem cells. Stem Cells Dev 18:661–670. https://doi.org/10.1089/scd.2008.0157
Dreyfuss JL, Regatieri CV, Jarrouge TR, Cavalheiro RP, Sampaio LO, Nader HB (2009) Heparan sulfate proteoglycans: structure, protein interactions and cell signaling. An Acad Bras Cienc 81:409–429
Gdalevitch M, Kasaai B, Alam N, Dohin B, Lauzier D, Hamdy RC (2013) The effect of heparan sulfate application on bone formation during distraction osteogenesis. PLoS ONE 8:e56790
Hausser HJ, Brenner RE (2004) Low doses and high doses of heparin have different effects on osteoblast-like Saos-2 cells in vitro. J Cell Biochem 91:1062–1073. https://doi.org/10.1002/jcb.20007
Irie A, Habuchi H, Kimata K, Sanai Y (2003) Heparan sulfate is required for bone morphogenetic protein-7 signaling. Biochem Biophys Res Commun 308:858–865. https://doi.org/10.1016/S0006-291X(03)01500-6
Kamoda H et al (2012) Platelet-rich plasma combined with hydroxyapatite for lumbar interbody fusion promoted bone formation and decreased an inflammatory pain neuropeptide in rats. Spine 37:1727–1733. https://doi.org/10.1097/BRS.0b013e31825567b7
Kasten P, Vogel J, Beyen I, Weiss S, Niemeyer P, Leo A, Lüginbuhl R (2008) Effect of platelet-rich plasma on the in vitro proliferation and osteogenic differentiation of human mesenchymal stem cells on distinct calcium phosphate scaffolds: the specific surface area makes a difference. J Biomater Appl 23:169–188. https://doi.org/10.1177/0885328207088269
Kim W, Jang CH, Kim G (2017) Optimally designed collagen/polycaprolactone biocomposites supplemented with controlled release of HA/TCP/rhBMP-2 and HA/TCP/PRP for hard tissue regeneration. Mater Sci Eng C 78:763–772
Latifi M, Talaei-Khozani T, Mehraban-Jahromi H, Sani M, Sadeghi-Atabadi M, Fazel-Anvari A, Kabir-Salmani M (2018) Fabrication of platelet-rich plasma heparin sulfate/hydroxyapatite/zirconia scaffold. Bioinspir Biomim Nanobiomater 7:122
Liao H-T, Chen J-P, Lee M-Y (2013) Bone tissue engineering with adipose-derived stem cells in bioactive composites of laser-sintered porous polycaprolactone scaffolds and platelet-rich plasma. Materials 6:4911–4929
Lin SS, Landesberg R, Chin HS, Lin J, Eisig SB, Lu HH (2006) Controlled release of PRP-derived growth factors promotes osteogenic differentiation of human mesenchymal stem cells. In: Engineering in Medicine and Biology Society, 2006. EMBS'06. 28th Annual International Conference of the IEEE, pp 4358–4361 https://doi.org/10.1109/IEMBS.2006.260847
M Dohan Ehrenfest D, Bielecki T, Jimbo R, Barbe G, Del Corso M, Inchingolo F, Sammartino G (2012) Do the fibrin architecture and leukocyte content influence the growth factor release of platelet concentrates? An evidence-based answer comparing a pure platelet-rich plasma (P-PRP) gel and a leukocyte-and platelet-rich fibrin (L-PRF). Curr Pharm Biotechnol 13:1145–1152. https://doi.org/10.2174/138920112800624382
Mansouri R et al (2017) Osteoblastic heparan sulfate glycosaminoglycans control bone remodeling by regulating wnt signaling and the crosstalk between bone surface and marrow cells. Cell Death Dis 8:e2902. https://doi.org/10.1038/cddis.2017.287
Mehdizadehkashi A et al (2017) Ultrastructural investigation of pelvic peritoneum in patients with chronic pelvic pain and subtle endometriosis in association with chromoendoscopy. J Minim Invasive Gynecol 24:114–123
Miao X, Hu Y, Liu J, Huang X (2007) Hydroxyapatite coating on porous zirconia. Mater Sci Eng C 27:257–261. https://doi.org/10.1016/j.msec.2006.03.009
Miguel BS, Kriauciunas R, Tosatti S, Ehrbar M, Ghayor C, Textor M, Weber FE (2010) Enhanced osteoblastic activity and bone regeneration using surface-modified porous bioactive glass scaffolds. J Biomed Mater Res Part A 94:1023–1033
Murali S, Manton KJ, Tjong V, Su X, Haupt LM, Cool SM, Nurcombe V (2009) Purification and characterization of heparan sulfate from human primary osteoblasts. J Cell Biochem 108:1132–1142. https://doi.org/10.1002/jcb.22340
Okuda K et al (2005) Platelet-rich plasma combined with a porous hydroxyapatite graft for the treatment of intrabony periodontal defects in humans: a comparative controlled clinical study. J Periodontol 76:890–898. https://doi.org/10.1902/jop.2005.76.6.890
Oryan A, Alidadi S, Moshiri A (2016) Platelet-rich plasma for bone healing and regeneration. Expert Opin Biol Ther 16:213–232. https://doi.org/10.1517/14712598.2016.1118458
Qi Y et al (2015) Combining mesenchymal stem cell sheets with platelet-rich plasma gel/calcium phosphate particles: a novel strategy to promote bone regeneration. Stem Cell Res Therapy 6:256. https://doi.org/10.1186/s13287-015-0256-1
Ren-fu Q et al (2012) Difference of adherence, proliferation and osteogenesis of mesenchymal stem cells cultured on different HA/ZrO2composites. Chin J Traumatol 15:131–139. https://doi.org/10.3760/cma.j.issn.1008-1275.2012.03.001
Rodriguez IA, Growney Kalaf EA, Bowlin GL, Sell SA (2014) Platelet-rich plasma in bone regeneration: engineering the delivery for improved clinical efficacy. BioMed Res Int. https://doi.org/10.1155/2014/392398
Sadeghi-Ataabadi M, Mostafavi-pour Z, Vojdani Z, Sani M, Latifi M, Talaei-Khozani T (2017) Fabrication and characterization of platelet-rich plasma scaffolds for tissue engineering applications. Mater Sci Eng C 71:372–380. https://doi.org/10.1016/j.msec.2016.10.001
Samuel S et al (2016) Platelet-rich concentrate in serum free medium enhances osteogenic differentiation of bone marrow-derived human mesenchymal stromal cells. PeerJ 4:e2347. https://doi.org/10.7717/peerj.2347
Sani F et al (2016) Differentiation of menstrual blood derived stem cell (MensSCs) to hepatocyte-liked cell on three dimensional nanofiberscaffold: poly caprolacton (PCL). J Biomed Sci Eng 9:216
Santo VE, Duarte ARC, Popa EG, Gomes ME, Mano JF, Reis RL (2012) Enhancement of osteogenic differentiation of human adipose derived stem cells by the controlled release of platelet lysates from hybrid scaffolds produced by supercritical fluid foaming. J Controll Release 162:19–27
Shadravanan M, Latifi M, Vojdani Z, Talaei-Khozani T (2020) Fabrication of Pentoxifylline-Loaded Hydroxyapatite/Alginate Scaffold for Bone Tissue Engineering. J Biomim Biomater Biomed Eng 47:25–40
Tanaka M et al (2017) Physico-chemical, in vitro, and in vivo evaluation of a 3D unidirectional porous hydroxyapatite scaffold for bone regeneration. Materials 10:33. https://doi.org/10.3390/ma10010033
Tang H-C, Chen W-C, Chiang C-W, Chen L-Y, Chang Y-C, Chen C-H (2015) Differentiation effects of platelet-rich plasma concentrations on synovial fluid mesenchymal stem cells from pigs cultivated in alginate complex hydrogel. Int J Mol Sci 16:18507–18521. https://doi.org/10.3390/ijms160818507
Uppada UK et al (2017) Combination of Bio-Gen mix®, platelet-rich fibrin and amnion membrane as a novel therapeutic option in regenerative periapical endodontic surgery: a case series. Int J Surg Case Rep. https://doi.org/10.1016/j.ijscr.2017.06.009
Vaishnavi C, Mohan B, Narayanan LL (2011) Treatment of endodontically induced periapical lesions using hydroxyapatite, platelet-rich plasma, and a combination of both: an in vivo study. J Conserv Dent 14:140
Yoshikawa H, Myoui A (2005) Bone tissue engineering with porous hydroxyapatite ceramics. J Artif Organs 8:131–136. https://doi.org/10.1007/s10047-005-0292-1
Acknowledgements
The authors wish to thank the council for stem cell science and technology and vice chancellery of research deputy of Shiraz University of medical Sciences for financial support.
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The authors wish to thank the council for stem cell science and technology and vice chancellery of research deputy of Shiraz University of medical Sciences for financial support (Grant No. 95-11-01-11880).
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All authors contributed to the study conception and design. An Adipose tissues collection and Mesenchymal Stem Cells isolation were done by Hanieh babakhanzadeh. Material preparation and analysis were performed by Mahin salmannejad and Mahsa Sani. Design of work, data collection and analysis were performed by Maryam kabir-salmani, Mona Latifi and Tahere Talaei-khozani. The first draft of the manuscript was written by Mona Latifi and edited by Tahere Talaei-khozani and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.
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Below is the link to the electronic supplementary material.Immunocytochemistry showed that cells on all scaffolds were stained for collagen I and all scaffolds had osteoinduction property; however, those cultured on HA/ZrO2/PRP/HS expressed more markers in the protein level
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latifi, M., Sani, M., Salmannejad, M. et al. Synergistic impact of platelet rich plasma-heparin sulfate with hydroxyapatite/zirconia on the osteoblast differentiation potential of adipose-derived mesenchymal stem cells. Cell Tissue Bank 23, 669–683 (2022). https://doi.org/10.1007/s10561-021-09966-0
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DOI: https://doi.org/10.1007/s10561-021-09966-0