IMMUNOHISTOCHEMICAL DETECTION OF GROWTH FACTORS AND EXTRACELLULAR MATRIX PROTEINS IN THE DEGENERATING TISSUES OF PRE-AND POSTNATAL HUMAN CLEFT LIP AND PALATE

태생 및 생후 구순.구개열에 나타나는 조직변성에 대한 성장인자와 세포외 기질 단백의 면역조직화학적 연구

  • Min, Bong-Gi (Department of Oral and Maxillofacial Surgery, College of Dentistry, Kangnung National University) ;
  • Lee, Suk-Keun (Department of Oral Pathology, College of Dentistry, Kangnung National University) ;
  • Park, Young-Wook (Department of Oral and Maxillofacial Surgery, College of Dentistry, Kangnung National University)
  • 민봉기 (강릉대학교 치과대학 구강악안면외과학교실) ;
  • 이석근 (강릉대학교 치과대학 구강병리학교실) ;
  • 박영욱 (강릉대학교 치과대학 구강악안면외과학교실)
  • Published : 2002.12.31

Abstract

In order to elucidate the pathogenesis of cleft lip and palate, first of all, it is necessary to understand the developmental mechanisms of growth factors and extracellular matrix proteins in the tissues of cleft lip and palate. We have performed immunohistochemical studies on human cleft lip and palate tissues to elucidate the pathogenetic implications of cleft lip and palate. 16 specimens from postnatal human cleft lip and palate subjects and 17 specimens from autopsy of prenatal human cleft lip and palate were fixed in 10% buffered formalin, embedded in paraffin. The sections were routinely stained by hematoxylin and eosin, also stained by PAS, and followed by immunohistochemical stainings using the antiseras of growth factors and extracellular matrix proteins such as PCNA, S-100, c-erb-B2, MMP-3, MMP-10, HSP-70, transglutaninase-C, E-cadherin, VEGF, vWF. Both the prenatal and postnatal specimens of cleft lip and palate showed dysplastic proliferation of the basal cell layer, increased infiltration of melanocytes into mucosal epithelium, sebaceous gland hyperplasia ingrowing into the muscular tissue of lip and palate, and fatty infiltration into the submucosal deep connective tissue. The strong reactions of MMP-3 and HSP-70 were detected in the tissues of cleft lip and palate, especially increased in degenerating muscle bundles, while the immunostainings of PCNA and c-erb-B2 were weakly positive in the tissues of cleft lip and palate. These data suggest that the retrogressive tissue degeneration around the cleft areas persistently exist during the prenatal and postnatal period after cleft formation, and the sebaceous gland hyperplasia and fatty infiltration with the intense expression of MMP-3 and HSP-70 is closely related to the muscular degeneration around the cleft area.

Keywords

References

  1. Kirschner RE, LaRossa D: Cleft lip and palate. Otolaryngol Clin North Am 2000;33:1191-1215 https://doi.org/10.1016/S0030-6665(05)70277-2
  2. Perrotin F, de Poncheville LM, Marret H, Paillet C, Lansac J, Body G: Chromosomal defects and associated malformations in fetal cleft lip with or without cleft palate. Eur J Obstet Gynecol Reprod Biol 2001;99:19-24 https://doi.org/10.1016/S0301-2115(01)00347-5
  3. Ferguson MW: Palate development. Development 1988;103:41-60
  4. Kaartinen V, Cui XM, Heisterkamp N, Groffen J, Shuler CF: Transforming growth factor-beta 3 regulates transdifferentiation of medial edge epithelium during palatal fusion and associated degradation of the basement membrane. Dev Dyn 1997;209:255-260 https://doi.org/10.1002/(SICI)1097-0177(199707)209:3<255::AID-AJA1>3.0.CO;2-H
  5. Shuler CF, Halpern DE, Guo Y, Sank AC: Medial edge epithelium fate traced by cell lineage analysis during epithelial-mesenchymal transformation in vivo. Dev Biol 1992;154:318-330 https://doi.org/10.1016/0012-1606(92)90071-N
  6. Bienengraber V, Malek F, Fanghanel J, Kundt G:Disturbances of palatogenesis and their prophylaxis in animal experiments. Anat Anz 1999;181:111-115 https://doi.org/10.1016/S0940-9602(99)80112-5
  7. Yano H, Yoshimoto H, Ohtsuru A, Ito M, Yamashita S, Fujii T: Characterization of cultured rat embryonic palatal mesenchymal cells. Cleft Palate Craniofac J 1996;33:379-384 https://doi.org/10.1597/1545-1569(1996)033<0379:COCREP>2.3.CO;2
  8. Lee SK, Kim YS, Jo YA, Seo JW, Chi JG: Prenatal development of cranial base in normal Korean fetuses. Anat Rec 1996;246:524-534 https://doi.org/10.1002/(SICI)1097-0185(199612)246:4<524::AID-AR11>3.0.CO;2-Q
  9. Lee SK, Kim YS, Lim CY, Chi JG: Prenatal growth pattern of the human maxilla. Acta Anat 1992;145:1-10 https://doi.org/10.1159/000147334
  10. Luning C, Rass A, Rozell B, Wroblewski J, Obrink B:Expression of E-cadherin during craniofacial development. J Craniofac Genet Dev Biol 1994;14:207-216
  11. Wang KY, Chang FH, Chiang CP, Chen KC, Kuo MYP:Temporal and spatial expression of erbB4 in ectodermal and mesenchymal cells during primary palatogenesis in noncleft and cleft strains of mice. J Oral Pathol Med 1998;27:141-146 https://doi.org/10.1111/j.1600-0714.1998.tb01930.x
  12. Martinez-Alvarez C, Bonelli R, Tudela C, Gato A, Mena J, O'Kane S, Ferguson MW: Bulging medial edge epithelial cells and palatal fusion. Int J Dev Biol 2000;44:331-335
  13. Blavier L, Lazaryev A, Groffen J, Heisterkamp N, DeClerck YA, Kaartinen V: TGF-beta3-induced palatogenesis requires matrix metalloproteinases. Mol Biol Cell 2001;12:1457-1466
  14. Mansell JP, Kerrigan J, McGill J, Bailey J, TeKoppele J, Sandy JR: Temporal changes in collagen composition and metabolism during rodent palatogenesis. Mech Ageing Dev 2000;119:49-62 https://doi.org/10.1016/S0047-6374(00)00168-8
  15. Morris-Wiman J, Du Y, Brinkley L: Occurrence and temporal variation in matrix metalloproteinases and their inhibitors during murine secondary palatal morphogenesis. J Craniofac Genet Dev Biol 1999;19:201-212
  16. Trotman CA, Hou D, Burdi AR, Cohen SR, Carlson DS:Histomorphologic analysis of the soft palate musculature in prenatal cleft and noncleft A/Jax mice. Cleft Palate Craniofac J 1995;32:455-462 https://doi.org/10.1597/1545-1569(1995)032<0455:HAOTSP>2.3.CO;2