猪胎儿釉基质衍生物刺激前成骨细胞2T9细胞增殖但不促进其分化，抑制成骨样MG63细胞增殖并促进其分化，还能增加正常人成骨细胞NHOst细胞的增殖和分化。

Porcine fetal enamel matrix derivative stimulates proliferation but not differentiation of pre-osteoblastic 2T9 cells, inhibits proliferation and stimulates differentiation of osteoblast-like MG63 cells, and increases proliferation and differentiation of normal human osteoblast NHOst cells.

作者信息

Schwartz Z, Carnes D L, Pulliam R, Lohmann C H, Sylvia V L, Liu Y, Dean D D, Cochran D L, Boyan B D

机构信息

Department of Orthopaedics, University of Texas Health Science Center, San Antonio 78229-3900, USA.

出版信息

J Periodontol. 2000 Aug;71(8):1287-96. doi: 10.1902/jop.2000.71.8.1287.

DOI:10.1902/jop.2000.71.8.1287

PMID:10972644

Abstract

BACKGROUND

Embryonic enamel matrix proteins are hypothesized to be involved in the formation of acellular cementum during tooth development, suggesting that these proteins can be used to regenerate periodontal tissues. Enamel matrix protein derived from embryonic porcine tooth germs is used clinically, but the mechanisms by which it promotes the formation of cementum, periodontal ligament, and bone are not well understood.

METHODS

This study examined the response of osteoblasts at 3 stages of osteogenic maturation to porcine fetal enamel matrix derivative (EMD). Proliferation (cell number and [3H]-thymidine incorporation), differentiation (alkaline phosphatase and osteocalcin), matrix synthesis ([35S]-sulfate incorporation; percentage of collagen production), and local factor production (prostaglandin E2 [PGE2] and transforming growth factor-beta 1 [TGF-beta1]) were measured in cultures of 2T9 cells (pre-osteoblasts which exhibit osteogenesis in response to bone morphogenetic protein-2 [BMP-2]), MG63 human osteoblast-like osteosarcoma cells, and normal human osteoblasts (NHOst cells).

RESULTS

EMD regulated osteoblast proliferation and differentiation, but the effects were cell-specific. In 2T9 cell cultures, EMD increased proliferation but had no effect on alkaline phosphatase-specific activity. EMD decreased proliferation of MG63 cells and increased cellular alkaline phosphatase and osteocalcin production. There was no effect on collagen synthesis, proteoglycan sulfation, or PGE2 production; however, TGF-beta1 content of the conditioned media was increased. There was a 60-fold increase in cell number in third passage NHOst cells cultured for 35 days in the presence of EMD. EMD also caused a biphasic increase in alkaline phosphatase that was maximal at day 14.

CONCLUSIONS

EMD affects early states of osteoblastic maturation by stimulating proliferation, but as cells mature in the lineage, EMD enhances differentiation.

摘要

背景

胚胎釉基质蛋白被认为在牙齿发育过程中参与无细胞牙骨质的形成，这表明这些蛋白可用于牙周组织再生。源自胚胎猪牙胚的釉基质蛋白已用于临床，但它促进牙骨质、牙周韧带和骨形成的机制尚不清楚。

方法

本研究检测了成骨成熟3个阶段的成骨细胞对猪胎儿釉基质衍生物（EMD）的反应。在2T9细胞（对骨形态发生蛋白-2 [BMP-2]有反应的前成骨细胞）、MG63人成骨样骨肉瘤细胞和正常人成骨细胞（NHOst细胞）培养物中测量增殖（细胞数量和[3H]-胸腺嘧啶核苷掺入）、分化（碱性磷酸酶和骨钙素）、基质合成（[35S]-硫酸盐掺入；胶原蛋白产生百分比）和局部因子产生（前列腺素E2 [PGE2]和转化生长因子-β1 [TGF-β1]）。

结果

EMD调节成骨细胞增殖和分化，但作用具有细胞特异性。在2T9细胞培养物中，EMD增加增殖，但对碱性磷酸酶特异性活性无影响。EMD降低MG63细胞的增殖并增加细胞碱性磷酸酶和骨钙素的产生。对胶原蛋白合成、蛋白聚糖硫酸化或PGE2产生无影响；然而，条件培养基中的TGF-β1含量增加。在存在EMD的情况下培养35天的第三代NHOst细胞中细胞数量增加了60倍。EMD还导致碱性磷酸酶呈双相增加，在第14天达到最大值。