Plastic and Reconstructive Surgery Research, Center for Dermatology Research, NIHR, Manchester Biomedical Research Center, University of Manchester, Manchester, United Kingdom.
Department of Biochemistry, Institute of Integrative Biology, University of Liverpool, Liverpool, United Kingdom.
Adv Wound Care (New Rochelle). 2021 Jan;10(1):1-12. doi: 10.1089/wound.2020.1154.
The goal of any topical formulation is efficient transdermal delivery of its active components. However, delivery of compounds can be problematic with penetration through tough layers of fibrotic dermal scar tissue. We propose a new approach combining high-performance liquid chromatography (HPLC) and Raman spectroscopy (RS) using a topical of unknown composition against a well-known antiscar topical (as control). Positive detection of compounds within the treatment topical using both techniques was validated with mass spectrometry. RS detected conformational structural changes; the 1,655/1,446 cm ratio estimating collagen content significantly decreased ( < 0.05) over weeks 4, 12, and 16 compared with day 0. The amide I band, known to represent collagen and protein in skin, shifted from 1,667 to 1,656 cm, which may represent a change from β-sheets in elastin to α-helices in collagen. Confirmatory elastin immunohistochemistry decreased compared with day 0, conversely the collagen I/III ratio increased in the same samples by week 12 ( < 0.05, and < 0.0001, respectively), in keeping with normal scar formation. Optical coherence tomography attenuation coefficient representing collagen deposition was significantly decreased at week 4 compared with day 0 and increased at week 16 ( < 0.05). This study provides a platform for further research on the simultaneous evaluation of the effects of compounds in cutaneous scarring by RS and HPLC, and identifies a role for RS in the therapeutic evaluation and theranostic management of skin scarring. RS can provide noninvasive information on the effects of topicals on scar pathogenesis and structural composition, validated by other analytical techniques.
任何局部制剂的目标都是高效地将其活性成分经皮递送至体内。然而,化合物的递送可能会成为问题,因为它们难以穿透纤维化的真皮瘢痕组织这一坚韧的层。我们提出了一种新方法,将高效液相色谱(HPLC)和拉曼光谱(RS)结合使用,对一种未知成分的局部制剂(作为治疗组)和一种已知的抗瘢痕局部制剂(作为对照组)进行检测。使用质谱法验证了这两种技术都能在治疗局部制剂中检测到化合物。RS 检测到了构象结构的变化;1655/1446 cm-1 比值估计胶原含量在第 4、12 和 16 周时与第 0 天相比显著降低( < 0.05)。酰胺 I 带,已知代表皮肤中的胶原和蛋白质,从 1667 cm-1 移至 1656 cm-1,这可能代表着从弹性蛋白中的β-折叠转变为胶原中的α-螺旋。与第 0 天相比,弹性蛋白免疫组织化学染色的确认减少,相反,在同一样本中,胶原 I/III 比值在第 12 周时增加( < 0.05 和 < 0.0001),与正常瘢痕形成一致。代表胶原沉积的光相干断层扫描衰减系数在第 4 周时与第 0 天相比显著降低,而在第 16 周时增加( < 0.05)。本研究为通过 RS 和 HPLC 同时评估化合物在皮肤瘢痕形成中的作用提供了一个平台,并确定了 RS 在皮肤瘢痕治疗评估和治疗管理中的作用。RS 可以提供关于局部制剂对瘢痕发病机制和结构组成影响的非侵入性信息,这一结果通过其他分析技术得到了验证。