Biocompatibility evaluation in vitro. Part I: Morphology expression and proliferation of human and rat osteoblasts on the biomaterials

Abstract

The biocompatibility evaluation of calcium phosphate based biomaterials is performed by tissue culture in vitro model. Three kinds of bioceramic materials which are potential to deal with bone trauma and/or conduct tissue growth are recommodated. The biological research results show that human and animal osteoblast cells anchor the materials surface in two hours in culture. Confocal laser scanning microscopy (CLSM) demonstrated the normal cell distribution and proliferation on both of dense and porous biomaterials. Hydroxyapatite and tricalcium phosphate stimulate cell proliferation. However, DNA and protein synthesis were considerably limited and the apoptosis phenomenon would be present on the hydroxyapatite (HA) materials by adding Al, Mg elements. Several important methods of biocompatibility evaluation of implant materials are described and the related biological molecular techniques such as tissue culture, cell transfection, cellular DNA stain, and Lowry assay are involved in the present research.

Keywords

biomaterials / tissue culture / cell morphology / proliferation

References

[1] BlackJBiological performance of materials: Fundamentals of biocompatibility[M], 1992, New York, Marcell Dekker Inc: 25-109

[2] BelloweC G, AubinJ E, HeerscheJ N M, et al.. Mineralised nodules formed in vitro from enzymatically released rat calvaria populations[J]. Calcif Tissue Int, 1986, 38: 143-154

[3] BiasiniV, MartinettiR, ManganoC, et al.. Interface between hydroxyapatite and mandibular human bone tissur[J]. Biomaterials, 1992, 13: 162-167

[4] AndersonH C. Mechanisms of mineral formation in bone[J]. Lab Invest, 1989, 60: 320-330

[5] AloiaJ F, VasmaniA, YehJ K, et al.. Calcitriol in the treatment of post menopausal osteoporosis[J]. Am J Med, 1988, 84: 401-408

[6] BenahmedM, BoulerJ M, GanO, et al.. Biodegradation of synthetic biphasic calcium phosphate by human monocytes in vitro: A morphological study[J]. Biomaterials, 1990, 11: 2173-2178

[7] DamienJ C, ParsonJ R. Bone grafts and bone graft substitutions[J]. J Appl Biomat, 1991, 2: 187-208

[8] GregoireM, OrlyI, MenanteauJ. The influence of calcium phosphate biomaterials on human bone cell activities: An in vitro approach[J]. J Biomed Mater Res, 1990, 24: 165-177

[9] JossetY, ZarrinpourA, LorenzatoM, et al.. Invitro reactions of human osteoblasts in culture with zirconia and alumina ceramics[J]. J Biomed Mater Res, 1999, 47: 481-493

[10] LaurenceiC T, AttawiaM A, ElgendyH M, et al.. Porous polymer-ceramic systems for tissue engineering support the formation of mineralised bone matrix[J]. Mat Res Soc Symp Proc, 1996, 414: 157-163

[11] MacnairR ABiocompatability study of orthopaedic materials with primary and immortalised osteoblast like cells derived from rat and human tissue[D], 1996, Glasgow, UK, Strathclyde University

[12] KushitaniS, IwakiH, SakaK, et al.WilsonJ, HenchL L, GreespanD, et al.. Comparative bone formation in several kinds of bioceramic granules[A]. Biomaterials[C], 1994, UK, Oxford, Alden Press: 137-144

[13] AndersonH C. Mechanisms of mineral formation in bone[J]. Lab Invest, 1989, 60: 320-330

[14] AndersonH C, MorrisD CMundyG R, MartinT J. Mineralisation[A]. Physiology and Pharmacology of Bone[C], 1993, London, Springer-Verlag: 267-298

[15] BagambisaF B, JoosU. Preliminary studies on the phenomenological behaviour of osteoblasts cultured on hydroxyapatite ceramics[J]. Biomaterials, 1990, 11: 50-56

[16] LowryO H, ResebroughN J, FarA L, et al.. Protein measurement with the folin phenol reagent[J]. J Biol Chem, 1951, 193: 265-275

[17] DoostdarH, DemozA, BurkeM D, et al.. Variation in drug-metabolising enzyme activities during the growth of human Hep G2 hepatoma cells[J]. Xenobiotica, 1990, 4: 435-441

[18] ZhenEZhenE, XiuJ C. Introduction of tissue culture[A]. Tissue culture and cytology, 1997, Beijing, Beijing Press: 3-39(in Chinese)

[19] AnselmeK, SharrockP, HadouinH, et al.. In vitro growth of human adult bone-derived cells on hydroxyapatite plasmasprayed coatings[J]. J Biomed Mater Res, 1996, 34: 247-259

[20] El-GhannamA, DucheyneP, ShapiroI M. Bioactive material template for in vitro synthesis of bone[J]. J Biomed Mater Res, 1995, 29: 359-370

[21] El-GlannamA, DucheyneP, ShapiroI M. Porous bioactive glass and hydroxyapatite ceramic affect bone cell function in vitro along different time lines[J]. J Biomed Mater Res, 1997, 36: 167-180

[22] SheppardC J R, ShottonD MConfocal laser scanning microscopy[M], 1997, Oxford, BIOS Scientific Publishers: 1-23

[23] Beltz B S and Burd G D. Immunocytochemical Techniques[M]. Blackwell Scientific Publications, 1989. 1–45.

[24] AdamsRlpLaboratory techniques in biochemistry an molecular biology[M], 1990, England, Oxford, Elsvier: 1-34

[25] FreshneyR ICulture of animal cells[M], 1994, England, London, Wiley-Liss Publication: 9-159

[26] LaceyA JLight microscopy in biology: a practical approach[M], 1989, New York, Oxford University Press: 9-24

[27] MacnairR A, WilkinsonR, MacDonaldC, et al.. Application of confocal laser scanning microscopy to cytocompatibility testing of potential orthopaedic materials in immortalised osteoblast-like cell lines[J]. Cells and Mater, 1996, 6: 71-78