Osteoarthritis is a prevalent degenerative joint disease characterized by progressive articular cartilage loss and destruction. The resultant increase in friction causes severe pain. The collagen I matrix (COL I) has been used clinically for cartilage repair; however, how COL I acts at cartilage surfaces is unclear. Here, we studied adsorption and lubrication of synovial fluid components, albumin, γ-globulin, and the phospholipid DPPC, on COL I under physiological conditions using surface plasmon resonance and an in situ sensing surface force apparatus. Our results revealed COL I had poor lubrication ability, a fairly high coefficient of friction (COF, μ = 0.651 ± 0.013), and surface damage under a 7 mN load. DPPC formed an improved lubricating layer on COL I (μ = 0.072 ± 0.016). In sharp contrast, albumin and γ-globulin exhibited poor lubrication with an order of magnitude higher COF but still provided benefits by protecting COL I from wear. Hence, DPPC on COL I may help optimize COL I implantation design.