Methylsulfonylmethane on Knee Laxity
Methylsulfonylmethane on Knee Laxity
The proposed research will focus on determining the effect of methylsulfonylmethane (MSM) on
 knee laxity changes through the menstrual cycle in young active females.

 As an extension to recent discovery, that MSM reverses the negative effect of estrogen on
 engineered ligament function, the proposed work is designed to determine whether MSM can
 decrease the negative effect of estrogen on knee laxity in females.

 Ligament function is determined by the content and cross-linking of collagen, which is
 influenced by a milieu of biochemical and mechanical parameters. The greater the amount and
 cross-linking the greater the stiffness and strength of these connective tissues. In
 engineered ligaments it has been previously shown that the high levels of estrogen, normally
 present in the days before and after ovulation, can inhibit the cross-linking enzyme lysyl
 oxidase. This decrease in collagen cross-linking likely increases connective tissue laxity
 and contributes to observed 4-fold greater occurrence of anterior cruciate ligament (ACL)
 rupture in females. Conversely, MSM increases collagen cross-linking and recent work
 conducted by the Baar lab in engineered human ligaments treated with high estrogen
 demonstrated that MSM could completely reverse the effects of estrogen on ligament mechanics.

 The proposed research aims to advance this promising pre-clinical data and apply in a
 clinical trial. This research also proposes to quantify that knee laxity increases up to 5mm
 between the first day of menstruation and the day after ovulation and also that the magnitude
 of the increase in laxity is directly related to the magnitude of the change in estrogen.
 Importantly, a direct relationship between knee laxity and ACL rupture exists. For every
 1.3mm increase in anterior-posterior knee displacement, the odds of ACL rupture increase
 4-fold. Therefore, any treatment that decreases knee laxity could be expected to reduce ACL
 ruptures and have widespread application across the general active population and high-level
 athletics.
Familiarization: Subjects would come to the Human Performance lab for a study orientation to
 be informed of the goals of the study, have anthropometric data collected (InBody770, In
 Body, Cerritos, CA) and familiarized with all testing and data collection tools. The purpose
 of this is to collect anthropometric data, review the study procedures and study
 requirements.

 Determination of menstruation date: After study familiarization instructions will be given to
 contact the research team and come back to the lab on the first day of menstruation to
 establish the individual study timeline. An ovulation kit will be provided. The ovulation kit
 would then be started at the end of the first reported menstrual cycle and be used daily
 until the 2-4 days of high estrogen (ovulation) are established.

 Baseline data: After menstruation date is established, for the first two months of the study,
 subjects would come to the lab on the first day of menstruation and the day after ovulation,
 have their knee laxity measured (GNRB dynamic laximeter, Prothia, Worcester, MA) and salivary
 analysis to determine estrogen levels. The first two months would determine normal changes in
 knee laxity and estrogen levels throughout the cycle within each subject.

 Intervention: Following the second ovulation testing day and once baseline values are
 established, subjects would be provided with 3 months of MSM or placebo control (rice flour)
 and instructed to take 2g each morning for the subsequent 3 months. Month 3 would serve as an
 MSM loading month and subjects would return to the laboratory throughout months 4 and 5 (day
 14 and first day of menstruation) for mechanical and estrogen testing.

 MSM or placebo treatments: The study will be a randomized double-blinded placebo-controlled
 trial. MSM or placebo capsules would be provided in a double blinded manner following the
 second mechanical test. Each individual would consume 2g of either the MSM or placebo control
 for the remainder of the study.

 Intervention Product and Preparation Intervention: MSM (OptiMSM®) is a pure, generally
 recognized as safe (GRAS) designated product. MSM can be found in small amounts in corn,
 tomatoes, tea, coffee, milk.

 Placebo: Rice flour is a commonly ingested food or additive. Intervention and placebo will be
 encapsulated in 1g vegetarian capsules (Hypromellose Capsule).

 Both intervention and placebo capsules are identically transparent with white powder inside.
 Capsules containers with 805 capsules (2g per day for each month plus extra) will be
 prepared, packaged by Bergstrom nutrition and transported with blinded code (A or B) so only
 individuals responsible for intervention and placebo preparation are removed from conducting
 the study and are off site (Vancouver, Washington).

 Product will be labelled by Bergstrom Nutrition. Subject ID and month (3, 4, or 5) will be
 filled in manually by University of California, Davis researcher upon randomization.

 Determination of knee laxity: Knee laxity will be determined using a GNRB dynamic laximeter
 following established protocols. Briefly, with the femur held in place, a controlled pressure
 will be applied to the calf. The pressure increases from 0 to 200 Newtons and a sensor
 measures kneecap displacement (laxity) in millimeters.

 Determination of estrogen levels: Salivary samples will be taken on the first day of
 menstruation and one day following ovulation for the 4 months of testing (month 1, 2, 4, 5).
 Estradiol levels in the saliva will be determined using the Estradiol Saliva ELISA kit (Rocky
 Mountain Diagnostics, CO) as per manufacturer's instructions.

 Data Collected:

 -  Record of menstrual cycle

 -  Salivary estradiol levels as described above

 -  Measurements of knee laxity
Inclusion Criteria:

 -  • Regular menstrual cycle (~1 x month)

 -  Not using oral contraceptive

 -  No history of serious knee injury (e.g. ACL rupture)

 -  Not currently pregnant (known)

 -  Not routinely taking medication or supplements that would interact with study
 measures or be contraindicated (e.g. steroid injections)

 Exclusion Criteria:

 -  • Irregular menstrual cycling (1 x month)

 -  Using oral contraceptive

 -  History of serious knee injury (e.g. ACL rupture)

 -  Currently pregnant or potential of being pregnant

 -  Routinely taking medication or supplements that would interact with study
 measures or be contraindicated (e.g. steroid injections)