Prolotherapy is a technique that involves the injection of an irritant, usually a hyperosmolar dextrose solution, typically in the treatment of chronic painful musculoskeletal conditions. Despite its long history and widespread use as a form of complementary therapy, there still are disparities over its optimal indications and injection preparations. There are, however, numerous studies available regarding the use and efficacy of prolotherapy for various musculoskeletal conditions. The most frequently published indication is in the treatment of chronic low back pain, but there are recent studies that examined its use in the management of refractory tendinopathies as well as osteoarthritis. There is growing evidence to suggest that prolotherapy may be helpful in treating chronic low back pain when coupled with adjunctive therapies, such as spinal manipulation or corticosteroid injections. There is also evidence to suggest that prolotherapy is effective in treating refractory tendinopathies, particularly for lateral epicondylosis and Achilles tendinopathy. Additional larger, randomized controlled trials are needed to make specific recommendations regarding ideal protocols and indications. There is emerging evidence for the use of prolotherapy as a treatment option for osteoarthritis; however, further studies are needed to conclusively demonstrate its efficacy. Overall, prolotherapy remains a promising option for the treatment of painful musculoskeletal conditions, particularly when other standard treatments have proved ineffective.