Rescue strategies by the topical application of TGFbeta3 and GDF5 to prevent a craniosynostosis in M. Crouzon mice and spatial evaluation of the gene expression pattern in the cranial vault and the dura mater of M. Crouzon mice.

颅缝闭锁的特点是颅缝过早闭合。这些是成纤维细胞生长因子受体2 （FGFR2）突变综合征的一部分，如Apert综合征和M. Crouzon，或零星发生。然而，大脑的自然生长受到影响，从而导致颅内压升高，导致神经元损伤。然后，手术分离这些缝合缝合线是必要的。然而，术后颅缝闭合复发的发生率为20%，需要进一步的手术干预。这项工作的目的是评估新的治疗方法，以防止小鼠术后颅缝闭锁复发。TGFbeta3已经在健康大鼠中成功应用。我们也将局部应用TGFbeta3和GDF5预防M. Crouzon术后颅缝紧闭复发。建立了几个实验组，以区分手术、支架（胶原）和生长因子对M.Crouzon和野生型小鼠颅缝闭锁复发的影响。通过超声和x线检查以及组织形态学和免疫组织学方法评估局部应用的成功。除了检查组织学参数外，还将对缝合线进行免疫组织化学检查。与先前发表的研究相反，脏器颅骨的缝合线也将包括在分析中。在另一系列的实验中，硬脑膜对颅缝的影响将被检查，因为它对颅缝紧闭的发展有旁分泌的影响。通过对硬脑膜和骨的蛋白表达进行地形图和三维可视化，比较野生型小鼠和M.Crouzon小鼠缝合线和硬脑膜的空间不匹配。这一证据也可能具有治疗意义。

Craniosynostoses are characterized by premature closure of cranial sutures. These occur as part of syndromes with Fibroblast Growth Factor Receptor 2 (FGFR2) mutations, e.g. the Apert syndrome and M. Crouzon, or occur sporadically. The natural growth of the brain is, however, affected and thus leads to an increase in intracranial pressure with the consequence of neuronal damage. Then, a surgical separation of these synostotic sutures is necessary. A postoperative recurrence of craniosynostosis happens, however, in 20% and demands a further surgical intervention. This work has the objective to evaluate new therapeutic approaches to prevent the recurrence of craniosynostoses postoperatively in mice suffering from M. Crouzon. In healthy rats TGFbeta3 has already been used successfully. We as well will apply TGFbeta3 and GDF5 locally to prevent a recurrence of craniosynostoses in M. Crouzon postoperatively. Several experimenal groups are established to differentiate the influence of surgery, scaffold (Collagen) and growth factors on the recurrence of craniosynostosis in M.Crouzon and wild type mice. The success of the local applications are evaluated by sonographic and x-ray examinations as well as histomorphologic and immunohistologic methods. In addition to the examination of histological parameters, the sutures will be also examined immunohistochemically. In contrast to previously published studies, the sutures of the viscerocranium will also be included in the analysis. In another series of experiments the influence of the dura mater on the cranial sutures will be examined, because is has a paracrine impact on the development of craniosynostoses. With the help of a topographic, 3D visualization of the protein expression of the dura mater and of the bone, wild-type mice and M.Crouzon mice will be compared in order to investigate spatial mismatches of sutures and dura mater. This proof could also have therapeutic implications.