Rastogi Mahima, Chowdhury Aniket, Chakraborty Sourabrata, Sahu Khageswar, Majumder Shovan Kumar
Laser Biomedical Applications Division, Raja Ramanna Centre for Advanced Technology, Indore, Madhya Pradesh 452013, India; Homi Bhabha National Institute, Training School Complex, Anushakti Nagar, Mumbai 400094, India.
Laser Biomedical Applications Division, Raja Ramanna Centre for Advanced Technology, Indore, Madhya Pradesh 452013, India; Homi Bhabha National Institute, Training School Complex, Anushakti Nagar, Mumbai 400094, India.
Spectrochim Acta A Mol Biomol Spectrosc. 2025 Mar 15;329:125552. doi: 10.1016/j.saa.2024.125552. Epub 2024 Dec 7.
Therapeutic applications involving mesenchymal stem cells (MSCs) encounter challenges of attaining therapeutically potent and scaled up number during in-vitro batch culture. Recently, photobiomodulation (PBM) has emerged as a non-pharmacological method for enhancing MSC number, potency, and secretome production. However, the absence of a versatile, non-invasive technique to accurately identify PBM-induced biochemical alterations hinders the clinical translation of the approach. Raman spectroscopy (RS) can be a potential solution to this challenge. In this study, we demonstrate the utility of micro-RS to determine red light (∼660 nm) induced molecular alterations in human adipose tissue derived MSCs (hADMSCs) exposed to three different red light (∼660 nm) fluence; ∼3, 6 and 9 J/cm. While the immediate changes in response to ∼660 nm exposure are subtle, at 6 and 24 h, there is increase in peak intensity of reducedCytochromes c, c1 and b, phenylalanine,CN stretching, CC lipids, OPO stretchingin cells. Maximum increase in intensity of these peaks was observed at ∼6 J/cm. Raman peak at 1585 cm, assigned to stretching vibration (CαCm) asymmetry of reduced Cyt c and sensitive to cellular redox status, shows notable change. Further, the intensity ratio of 1585 cm and 1452 cm, a suggestive Raman biomarker for cell proliferation, is increased in cells exposed to ∼3 & ∼6 J/cm followed by a decrease in cells exposed to ∼9 J/cm. Furthermore, both micro-RS intensity ratio (1585 cm/1452 cm) and MTT data on cell viability are in qualitative agreement with each other and show biphasic response to ∼660 nm exposure. While these results suggest the utility of micro-RS for label free assessment of PBM induced changes in hADMSCs, detailed studies on other cell types are necessary to validate the utility of micro-RS in this field.
涉及间充质干细胞（MSC）的治疗应用在体外批量培养过程中面临着获得具有治疗效力且数量可扩大的挑战。最近，光生物调节（PBM）已成为一种用于增加MSC数量、效力和分泌组产生的非药物方法。然而，缺乏一种通用的、非侵入性技术来准确识别PBM诱导的生化改变阻碍了该方法的临床转化。拉曼光谱（RS）可能是应对这一挑战的潜在解决方案。在本研究中，我们展示了显微拉曼光谱在确定红光（660nm）诱导的人脂肪组织来源的间充质干细胞（hADMSC）分子变化中的应用，这些细胞暴露于三种不同的红光（660nm）通量；3、6和9J/cm²。虽然对660nm暴露的即时反应变化细微，但在6小时和24小时时，细胞中还原型细胞色素c、c1和b、苯丙氨酸、C≡N伸缩振动、C-C脂质、P-O-P伸缩振动的峰强度增加。在6J/cm²处观察到这些峰强度的最大增加。在1585cm⁻¹处的拉曼峰，归因于还原型细胞色素c的伸展振动（Cα-Cm）不对称性且对细胞氧化还原状态敏感，显示出显著变化。此外，1585cm⁻¹和1452cm⁻¹的强度比，这是一种提示细胞增殖的拉曼生物标志物，在暴露于3和6J/cm²的细胞中增加，随后在暴露于9J/cm²的细胞中降低。此外，显微拉曼光谱强度比（1585cm⁻¹/1452cm⁻¹）和关于细胞活力的MTT数据在质量上彼此一致，并显示出对~660nm暴露的双相反应。虽然这些结果表明显微拉曼光谱可用于无标记评估PBM诱导的hADMSC变化，但有必要对其他细胞类型进行详细研究以验证显微拉曼光谱在该领域的实用性。
