title: Topical application of laminin-332 to diabetic mouse wounds.
abstract: BACKGROUND
  Keratinocyte migration is essential for wound healing and diabetic wound keratinocytes migrate poorly. Keratinocyte migration and anchorage appears to be mediated by laminin-332 (LM-332). Impaired diabetic wound healing may be due to defective LM-332 mediated keratinocyte migration.
  
  
  OBJECTIVE
  To evaluate LM-332 expression in diabetic (db/db) and control (db/-) mice and to test LM-332 wound healing effects when applied to mouse wounds.
  
  
  METHODS
  LM-332 expression in mouse wounds was evaluated using immunohistochemistry. LM-332 wound healing effects were evaluated by directly applying soluble LM-332, a LM-332 biomaterial, or a control to mouse wounds. Percent wound closure and histology score, based on healing extent, were measured.
  
  
  RESULTS
  Precursor LM-332 expression was markedly reduced in db/db when compared to db/- mice. In vitro, soluble LM-332 and LM-332 biomaterial demonstrated significant keratinocyte adhesion. In vivo, soluble LM-332 treated wounds had the highest histology score, but significant differences were not found between wound treatments (p>0.05). No differences in percentage wound closure between treatment and control wounds were found (p>0.05).
  
  
  CONCLUSIONS
  The db/db wounds express less precursor LM-332 when compared to db/-. However, LM-332 application did not improve db/db wound healing. LM-332 purified from keratinocytes was primarily physiologically cleaved LM-332 and may not regulate keratinocyte migration. Application of precursor LM-332 rather than cleaved LM-332 may be necessary to improve wound healing, but this isoform is not currently available in quantities sufficient for testing.
authors: Department of Surgery (Plastic and Reconstructive Surgery), University of Washington, Seattle, WA 98195-6524, United States.
authors: Stephen R
authors: SR
authors: Sullivan
authors: Robert A
authors: RA
authors: Underwood
authors: Randall O
authors: RO
authors: Sigle
authors: Yuko
authors: Y
authors: Fukano
authors: Lara A
authors: LA
authors: Muffley
authors: Marcia L
authors: ML
authors: Usui
authors: Nicole S
authors: NS
authors: Gibran
authors: Marcos A
authors: MA
authors: Antezana
authors: William G
authors: WG
authors: Carter
authors: John E
authors: JE
authors: Olerud
mesh_terms: D000287:Administration, Topical; D000818:Animals; D001485:Basement Membrane / Q000378:metabolism / Q000473:pathology; D015815:Cell Adhesion Molecules / Q000008:administration & dosage / Q000378:metabolism / Q000627:therapeutic use*; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology; D003094:Collagen / Q000378:metabolism; D048909:Diabetes Complications / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D016023:Integrins / Q000378:metabolism; D015603:Keratinocytes / Q000378:metabolism / Q000473:pathology / Q000502:physiology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D016688:Mice, Inbred NOD; D008817:Mice, Mutant Strains; D014945:Wound Healing / Q000187:drug effects; D014947:Wounds and Injuries / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D000097794:Kalinin
chemical_list: D015815:Cell Adhesion Molecules; D003094:Collagen; D016023:Integrins; D000097794:Kalinin
references: J Biol Chem. 2001 Aug 31;276(35):33045-53
references: 11395486
references: J Cell Sci. 2004 Sep 1;117(Pt 19):4481-94
references: 15316072
references: J Invest Dermatol. 1994 Nov;103(5 Suppl):31S-38S
references: 7963682
references: J Invest Dermatol. 2002 Dec;119(6):1400-4
references: 12485446
references: J Cell Biol. 1990 Dec;111(6 Pt 2):3141-54
references: 2269668
references: Plast Reconstr Surg. 2004 Mar;113(3):953-60
references: 15108888
references: Wound Repair Regen. 2002 Sep-Oct;10(5):295-301
references: 12406165
references: J Cell Sci. 2004 Mar 15;117(Pt 8):1351-63
references: 14996912
references: EMBO J. 2002 Aug 1;21(15):3919-26
references: 12145193
references: J Cell Sci. 1996 Oct;109 ( Pt 10):2509-20
references: 8923212
references: Exp Cell Res. 1996 Sep 15;227(2):309-22
references: 8831569
references: Diabet Med. 1996;13 Suppl 1:S6-11
references: 8741821
references: J Cell Biol. 1996 Feb;132(4):727-40
references: 8647901
references: Diabetes. 1983 Nov;32(11):1048-54
references: 6357903
references: Clin Exp Dermatol. 1996 Jan;21(1):23-7
references: 8689764
references: Acta Derm Venereol. 1995 Nov;75(6):459-62
references: 8651025
references: J Cell Biol. 1992 Nov;119(3):695-703
references: 1383241
references: J Cell Sci. 1999 Aug;112 ( Pt 16):2615-29
references: 10413670
references: J Dermatol. 1992 Nov;19(11):673-5
references: 1293153
references: J Burn Care Rehabil. 1999 Mar-Apr;20(2):155-62
references: 10188114
references: Cell. 1991 May 17;65(4):599-610
references: 2032285
references: Eur J Biochem. 1994 Jan 15;219(1-2):209-18
references: 8306988
references: Differentiation. 1984;26(1):49-54
references: 6370774
references: Exp Cell Res. 1997 Jun 15;233(2):330-9
references: 9194495
references: J Biol Chem. 1995 Oct 6;270(40):23496-503
references: 7559513
references: Diabetes. 1982;31(Suppl 1 Pt 2):1-6
references: 7160533
references: Methods Cell Biol. 2002;69:27-52
references: 12070998
references: Am J Med. 1981 Feb;70(2):353-60
references: 6451172
references: J Invest Dermatol. 1983 Jul;81(1 Suppl):33s-40s
references: 6345690
references: Diabetes Care. 1999 Aug;22(8):1354-60
references: 10480782
references: J Cell Biol. 1991 Aug;114(3):567-76
references: 1860885
references: Matrix Biol. 1994 Apr;14(3):209-11
references: 7921537
references: J Cell Biol. 1999 Jun 14;145(6):1309-23
references: 10366601
references: Diabetes Care. 1990 May;13(5):513-21
references: 2351029
references: J Biol Chem. 2001 Nov 23;276(47):43860-70
references: 11571278
references: Dev Cell. 2003 Aug;5(2):257-71
references: 12919677
references: J Cell Biol. 2001 Apr 30;153(3):465-78
references: 11331299
references: Endocrinology. 2000 Jan;141(1):446-9
references: 10614668
references: Diabetologia. 1978 Mar;14(3):141-8
references: 350680
references: J Cell Biol. 1998 Apr 6;141(1):255-65
references: 9531563
references: Wound Repair Regen. 1999 Mar-Apr;7(2):119-26
references: 10231513
references: J Biol Chem. 2000 Oct 13;275(41):31896-907
references: 10926936
references: Am J Pathol. 1990 Jun;136(6):1235-46
references: 2356856