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Planta Med 2024; 90(02): 96-110
DOI: 10.1055/a-2192-2138
Biological and Pharmacological Activity
Original Papers

Bone Fracture-healing Properties and UPLC-MS Analysis of an Enriched Flavonoid Fraction from Oxystelma esculentum

Priyanka Rawat‡
1   Sophisticated Analytical Instrument Facility & Research, CSIR-Central Drug Research Institute, Lucknow, Uttar Pradesh, India
3   Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, Uttar Pradesh, India
,
Kriti Sharma‡
2   Division of Endocrinology, CSIR-Central Drug Research Institute, Lucknow, Uttar Pradesh, India
3   Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, Uttar Pradesh, India
,
Sonu Khanka
2   Division of Endocrinology, CSIR-Central Drug Research Institute, Lucknow, Uttar Pradesh, India
3   Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, Uttar Pradesh, India
,
Yatendra Singh
1   Sophisticated Analytical Instrument Facility & Research, CSIR-Central Drug Research Institute, Lucknow, Uttar Pradesh, India
,
Ravi Prakash
2   Division of Endocrinology, CSIR-Central Drug Research Institute, Lucknow, Uttar Pradesh, India
,
Nisha Rais
1   Sophisticated Analytical Instrument Facility & Research, CSIR-Central Drug Research Institute, Lucknow, Uttar Pradesh, India
,
Arvind Kumar Maurya
1   Sophisticated Analytical Instrument Facility & Research, CSIR-Central Drug Research Institute, Lucknow, Uttar Pradesh, India
3   Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, Uttar Pradesh, India
,
Ritika Gupta
2   Division of Endocrinology, CSIR-Central Drug Research Institute, Lucknow, Uttar Pradesh, India
,
Dipak K. Mishra
1   Sophisticated Analytical Instrument Facility & Research, CSIR-Central Drug Research Institute, Lucknow, Uttar Pradesh, India
3   Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, Uttar Pradesh, India
,
Divya Singh
2   Division of Endocrinology, CSIR-Central Drug Research Institute, Lucknow, Uttar Pradesh, India
3   Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, Uttar Pradesh, India
,
Sanjeev Kanojiya 
1   Sophisticated Analytical Instrument Facility & Research, CSIR-Central Drug Research Institute, Lucknow, Uttar Pradesh, India
3   Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, Uttar Pradesh, India
› Author AffiliationsThis study was funded by the Council of Scientific and Industrial Research (CSIR), New Delhi under projects CII7041, MLP2028, and MLP2035. The CSIR-CDRI communication number is 10692.
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Abstract

Oxystelma esculentum has been used as a folk medicine to treat jaundice, throat infections, and skin problems. In the current study, the bone fracture-healing properties of a flavonoid-enriched fraction (Oxy50-60F) of O. esculentum were investigated in Swiss mice using a drill-hole injury model. Oxy50-60F (1 mg/kg/day, 5 mg/kg/day, and 10 mg/kg/day) was administered orally (from the next day) after a 0.6 mm drill-hole injury in mice femur mid-diaphysis for 7 days and 14 days. Parathyroid hormone (40 µg/kg; 5 times/week) was given subcutaneously as the positive control. Confocal imaging for bone regeneration, micro-architecture of femur bones, ex vivo mineralization, hematoxyline and eosin staining, measurement of reactive oxygen species, and gene expression of osteogenic and anti-inflammatory genes were studied. Quercetin, kaempferol, and isorhamnetin glycosides were identified in the active fraction using mass spectrometry techniques. Our results confirm that Oxy50-60F treatment promotes fracture healing and callus formation at drill-hole sites and stimulates osteogenic and anti-inflammatory genes. Oxy50-60F administration to fractured mice exhibited significantly better micro-CT parameters in a dose-dependent manner and promoted nodule mineralization at days 7 and 14 post-injury. Oxy50-60F also prevents ROS generation by increasing expression of the SOD2 enzyme. Overall, this study reveals that Oxy50-60F has bone regeneration potential in a cortical bone defect model, which supports its use in delayed-union and non-union fracture cases.

Key words

Oxystelma esculentum - Asclepiadaceae - flavonoids - fracture healing - osteoblast differentiation - drill-hole injury

Supporting Information

    An overlay MRM chromatogram of reference standards and Oxy50-60F (Fig. 1S), the LC-MS/MS data of all 17 compounds and standards (Fig. 2S–23S), the MRM chromatograms of standards for quantitative analysis (Fig. 24S), the calibration curves of standards (Fig. 25S–28S), a bar graph of the relative quantity of identified flavonoids in Oxy50-60F (Fig. 29S), and a table of linear-gradient for qualitative and quantitative analysis (Tables 1S and 2S) are available as Supporting Information.

  • Supporting Information


Publication History

Received: 25 September 2022

Accepted after revision: 16 October 2023

Accepted Manuscript online:
16 October 2023

Article published online:
10 November 2023

© 2023. Thieme. All rights reserved.

Georg Thieme Verlag KG
Rüdigerstraße 14, 70469 Stuttgart, Germany

 

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