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Histologic and Competitive Reverse-Transcription–Polymerase Chain Reaction Studies

Liancun Wu, MD; Aamir Siddiqui, MD; David E. Morris, MD; David A. Cox, PhD; Sanford I. Roth, MD; Thomas A. Mustoe, MD

Arch Surg. 1997;132(7):753-760.

Abstract
Background
Transforming growth factor (TGF) β3 is a new isoform of the TGFβ superfamily and is presumed to play an important role in wound repair and scarring.

Objective
To examine the effects of TGFβ3 on wound healing and on reducing scarring.

Design and Interventions
Dermal ulcers were created on the ears of 75 anesthetized young female rabbits. Either TGFβ3 or vehicle was applied topically to the wounds. Wounds were bisected and analyzed histologically at postwounding day 7. A second group of wounds was treated with topical TGFβ3 and TGFβ2 or vehicle at days 0 and 3 and harvested at days 21 through 42 as an excessive scarring model. The third group of wounds was treated with TGFβ1, TGFβ2, and TGFβ3 and vehicle. The granulation tissue was harvested at day 7, and cellular RNA was extracted for performing competitive reverse-transcription polymerase chain reaction.

Main Outcome Measurement
The amount of new epithelium and granulation tissue was measured in TGFβ3- and vehicle-treated wounds. The hypertrophic index was calculated for scarring wounds treated with TGFβ2 and TGFβ3 or vehicle. Levels of TGFβ1 messenger RNA were measured in those wounds that were treated with TGFβ1, TGFβ2, and TGFβ3 and in their controls.

Results
The use of TGFβ3 (0.3-0.75) µg per wound) increased granulation tissue formation by more than 100% (P<.005). Epithelialization showed a biphase, either increasing 30% (P<.04) or decreasing 25% (P<.001) dependent on dose. No significant difference in the hypertrophic index was noted in TGFβ3-treated wounds compared with controls. Levels of TGFβ1 messenger RNA increased (7.1- to 14.9-fold) in those wounds treated with TGFβ2 compared with controls at day 7.

Conclusions
Exogenous TGFβ3 displays substantial vulnerary properties in wound healing and may be useful in treating nonhealing wounds. However, the observation that TGFβ3 can reduce scarring was not confirmed in this study, and the messenger RNA level in response to TGFβ3 suggests that it behaves similarly to TGFβ1.

Arch Surg. 1997;132:753-760

Author Affiliations
From the Division of Plastic and Reconstructive Surgery (Drs Wu, Siddiqui, Morris, and Mustoe) and Department of Pathology (Dr Roth), Northwestern University Medical School, Chicago, Ill, and the Pharmaceutical Division, Molecular Biology Resources, Ciba-Geigy Ltd, Basel, Switzerland (Dr Cox). Dr Siddiqui is now with the Division of Plastic and Reconstructive Surgery, University of Minnesota Medical School, Minneapolis, and Dr Morris is with the Department of Surgery, Henry Ford Hospital, Detroit, Mich.

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