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For in vivo repair studies, histological, biomechanical, and functional endpoints should be evaluated alongside molecular mechanism markers to provide comprehensive characterisation of combination effects at tissue and molecular levels simultaneously

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You would need to draw twice, introducing additional error and contamination risk with each needle puncture

Methotrexate is given at high, potentially toxic doses followed by leucovorin rescue, and high drug levels can result in acute kidney injury (AKI)

doi: 10.2174/138920371604150429154840 97 ErdamarHCimenBGlcemalHSaraymenRYererBDemirciH

Cucinotta D, Vanelli M