Navia J M, Snider C, Punyasingh J, Harris S S
Arch Oral Biol. 1984;29(11):911-20. doi: 10.1016/0003-9969(84)90091-8.
A culture procedure for rat third molars suitable for nutritional-developmental studies is described. Unerupted third molars from 12-day-old rats were cultured in BGJb media containing 20 per cent rat serum and supplemented with 25 mM HEPES buffer, 25 mg ascorbic acid, 20 mg L-glutamine, 12 mg penicillin G and 10 mg streptomycin sulphate per 100 ml of media. Molars were cultured at the liquid-gas interphase using a 50 per cent O2, 45 per cent N2, 5 per cent CO2 gas mixture at 10 lb-psig (pounds per square inch guage). Molar cultures were maintained successfully for 9-14 days without evidence of necrosis, although they developed at a slower rate than in vivo. Molars cultured in 50 per cent O2 compared to those cultured in 21 per cent O2 for periods of 2, 4, 6 and 8 days had higher values for protein, alkaline phosphatase (AP), Ca, P and Ca/P. Vitamin-A-deficiency gave lower values for AP, Ca, P, Ca/P, 45Ca, 35S and [14C]-proline uptake. Histologically, A - molars had atrophic ameloblasts, some foci of squamous metaplasia and abnormal keratin formation. Thus, deficiency of vitamin A imposed during in-vitro development of rat third molars retarded dentinogenesis and interfered with early mineralization of enamel and dentine.
本文描述了一种适用于营养发育研究的大鼠第三磨牙培养方法。将12日龄大鼠未萌出的第三磨牙培养于含有20%大鼠血清的BGJb培养基中，每100 ml培养基补充25 mM HEPES缓冲液、25 mg抗坏血酸、20 mg L-谷氨酰胺、12 mg青霉素G和10 mg硫酸链霉素。磨牙在液-气界面培养，使用50% O₂、45% N₂、5% CO₂的气体混合物，压力为10磅/平方英寸（表压）。磨牙培养物成功维持9 - 14天，无坏死迹象，尽管其发育速度比体内慢。与在21% O₂中培养2、4、6和8天的磨牙相比，在50% O₂中培养的磨牙蛋白质、碱性磷酸酶（AP）、钙、磷和钙/磷值更高。维生素A缺乏导致AP、钙、磷、钙/磷、⁴⁵Ca、³⁵S和[¹⁴C]-脯氨酸摄取值降低。组织学上，维生素A缺乏的磨牙有成釉细胞萎缩、一些鳞状化生灶和异常角蛋白形成。因此，在大鼠第三磨牙体外发育过程中缺乏维生素A会延迟牙本质形成，并干扰釉质和牙本质的早期矿化。
