Photodynamic therapy on keloid fibroblasts in tissue-engineered keratinocyte-fibroblast co-culture

被引:58
作者
Chiu, LL
Sun, CH
Yeh, AT
Torkian, B
Karamzadeh, A
Tromberg, B
Wong, BJF [1 ]
机构
[1] Univ Calif Irvine, Beckman Laser Inst, Irvine, CA 92612 USA
[2] Univ Calif Irvine, Dept Otolaryngol Head & Neck Surg, Irvine, CA USA
[3] Univ Calif Irvine, Dept Biomed Engn, Irvine, CA USA
[4] Texas A&M Univ, Dept Biomed Engn, College Stn, TX USA
关键词
keloid; photodynamic therapy; PDT; raft; tissue engineering; multiphoton microscopy; two-photon microscopy; second harmonic generation; skin; neck; lasers; wound healing;
D O I
10.1002/lsm.20213
中图分类号
R75 [皮肤病学与性病学];
学科分类号
100206 ;
摘要
Background and Objectives: Keloids are disfiguring, proliferative scars that are a pathologic response to cutaneous injury. An organotypic tissue culture system (the Raft model [1-10]) was used to investigate the feasibility of using photodynamic therapy (PDT) as an adjunctive therapy to treat keloids following surgical excision. The Raft co-culture system mimics skin by layering keratinocytes on top of fibroblasts embedded in a collagen matrix. PDT uses drugs that produce singlet oxygen in situ when irradiated by light, and may lead to a number of effects in living tissues varying from the modulation of growth to apoptosis. PDT is already used to treat several benign and malignant diseases in organs such as the skin, retina, and esophagus. Study Design/Materials and Methods: Normal adult, neonatal, and keloid fibroblasts and keratinocytes were isolated from skin obtained from patients undergoing elective procedures and used to construct the Rafts. Mature Rafts (after 4 days) were incubated with 5-amino levulinic acid (5-AlA), a photosensitizer, for 3 hours and were laser-irradiated (635 nm) for total energy delivery of 5 J/cm(2), 10 J/cm(2), or 20 J/cm(2). Rafts were examined 24 hours and 14 days later. Cell viability was determined using confocal imaging combined with live-dead fluorescent dyes. Multiphoton microscope (MPM) imaged collagen structure and density. As Rafts contract over time, surface area was measured using optical micrometry daily. Results: At 10 and 20 J/cm(2), near-total cell death was observed in all constructs, while at 5 J/cm(2) cell viability was comparable to controls. Cell viability in keloid and neonatal Rafts was greater than that observed in normal adult Rafts. Treated Rafts contracted less over the 14-day period compared to controls. Contraction and collagen density were greatest in keloid and neonatal Rafts. Conclusions: A PDT dosimetry range was established, which reduces tissue contraction and collagen density while minimizing injury to fibroblasts.
引用
收藏
页码:231 / 244
页数:14
相关论文
共 67 条
[51]   Antivaccinia activities of acyclic nucleoside phosphonate derivatives in epithelial cells and organotypic cultures [J].
Snoeck, R ;
Holy, A ;
Dewolf-Peeters, C ;
Van Den Oord, J ;
De Clercq, E ;
Andrei, G .
ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, 2002, 46 (11) :3356-3361
[52]   Interleukin-1-stimulated secretion of interleukin-8 and growth-related oncogene-α demonstrates greatly enhanced keratinocyte growth in human raft cultured epidermis [J].
Steude, J ;
Kulke, R ;
Christophers, E .
JOURNAL OF INVESTIGATIVE DERMATOLOGY, 2002, 119 (06) :1254-1260
[53]   Alterations in fibroblast α1β1 integrin collagen receptor expression in keloids and hypertrophic scars [J].
Szulgit, G ;
Rudolph, R ;
Wandel, A ;
Tenenhaus, M ;
Panos, R ;
Gardner, H .
JOURNAL OF INVESTIGATIVE DERMATOLOGY, 2002, 118 (03) :409-415
[54]   Expression of Bcl-2, p53, c-jun and c-fos protooncogenes in keloids and hypertrophic scars [J].
Teofoli, P ;
Barduagni, S ;
Ribuffo, M ;
Campanella, A ;
De Pita, O ;
Puddu, P .
JOURNAL OF DERMATOLOGICAL SCIENCE, 1999, 22 (01) :31-37
[55]  
Tilkorn H, 1994, Polim Med, V24, P31
[56]  
Torkian Behrooz A, 2004, Arch Facial Plast Surg, V6, P180, DOI 10.1001/archfaci.6.3.180
[57]   A trial of short incubation, broad-area photodynamic therapy for facial actinic keratoses and diffuse photodamage [J].
Touma, D ;
Yaar, M ;
Whitehead, S ;
Konnikov, N ;
Gilchrest, BA .
ARCHIVES OF DERMATOLOGY, 2004, 140 (01) :33-40
[58]   Increased plasminogen activator inhibitor-1 in keloid fibroblasts may account for their elevated collagen accumulation in fibrin gel cultures [J].
Tuan, TL ;
Wu, HY ;
Huang, EY ;
Chong, SSN ;
Laug, W ;
Messadi, D ;
Kelly, P ;
Le, A .
AMERICAN JOURNAL OF PATHOLOGY, 2003, 162 (05) :1579-1589
[59]   ALTERED STEADY-STATE RATIO OF TYPE-I/III PROCOLLAGEN MESSENGER-RNAS CORRELATES WITH SELECTIVELY INCREASED TYPE-I PROCOLLAGEN BIOSYNTHESIS IN CULTURED KELOID FIBROBLASTS [J].
UITTO, J ;
PEREJDA, AJ ;
ABERGEL, RP ;
CHU, ML ;
RAMIREZ, F .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1985, 82 (17) :5935-5939
[60]   The effects of a single dose of 5-fluorouracil on keloid scars: A clinical trial of timed wound irrigation after extralesional excision [J].
Uppal, RS ;
Khan, U ;
Kakar, S ;
Talas, G ;
Chapman, P ;
McGrouther, AD .
PLASTIC AND RECONSTRUCTIVE SURGERY, 2001, 108 (05) :1218-1224